ARTICLE |
Correspondence to: Meelis Kolmer, Dept. of Human Molecular Genetics, National Public Health Institute, Mannerheimintie 166, FIN-00300 Helsinki, Finland. E-mail: meelis.kolmer@ktl.fi
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Summary |
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Autoimmune polyendocrinopathy candidiasis ectodermal dystrophy (APECED), also known as autoimmune polyglandular syndrome Type I (APS1), is an autosomal recessive autoimmune disease caused by mutations in a gene designated as AIRE (autoimmune regulator). Here we have studied the expression of Aire in transfected cell lines and in adult mouse tissues. Our results show that Aire has a dual subcellular location and that it is expressed in multiple immunologically relevant tissues such as the thymus, spleen, lymph nodes, and bone marrow. In addition, Aire expression was detected in various other tissues such as kidney, testis, adrenal glands, liver, and ovary. These findings suggest that APECED protein might also have a function(s) outside the immune system.
(J Histochem Cytochem 49:197208, 2001)
Key Words: APECED, APS1, PGA I, AIRE, autoimmunity, gene, expression, disease
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Introduction |
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AUTOIMMUNE polyendocrinopathy candidiasis ectodermal dystrophy (APECED; OMIM accession number 240300), also known as autoimmune polyglandular syndrome Type I (APS1), is a monogenic autosomal recessive autoimmune disease that affects several different organs. APECED symptoms fall into three main categories: (a) autoimmune polyendocrinopathy, (b) chronic mucocutaneous candidiasis, and (c) ectodermal dystrophies. A gene designated AIRE, defective in APECED patients has been identified (
The AIRE gene encodes a 58-kD protein containing a nuclear location signal and several structural domains characteristic of transcription regulators (
In transient expression studies using mammalian tissue culture cell lines, dual subcellular localization of the APECED protein, nuclear and cytoplasmic, has been observed (
On the basis of structural similarity to other proteins with an established role in the regulation of transcription and its subcellular location, APECED protein has been suggested to be a transcriptional regulator. Our recent results show that the human APECED protein is indeed able to promote transcription from the minimal promoter in mammalian one-hybrid assays (
Although the biological function(s) of the APECED protein have just began to emerge, the molecular mechanisms triggered by the mutations in the AIRE gene that finally result in the development of APECED, characterized by such a broad spectrum of clinical symptoms, remain elusive. Defining the biological functions of the AIRE and eventually understanding the pathogenesis of APECED is intimately bound to knowledge about where and when the AIRE gene is expressed.
In human, the expression of the APECED protein has been characterized in immunologically relevant tissues and was shown to be restricted to limited cell populations (
To extend our knowledge of the biological function of Aire gene, we explored its expression in transfected cell lines and in different mouse tissues using several different experimental approaches, i.e., RT-PCR, in situ mRNA hybridization, and immunohistochemical staining.
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Materials and Methods |
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cDNA Library Screening
An oligo(dT)-primed ZAPII 17d mouse kidney library (kindly provided by Dr. V. Olkkonen; National Public Health Institute, Helsinki, Finland) was screened using human AIRE full-length cDNA as a probe. The positive clones were plaque-purified through three subsequent screening cycles, subcloned into the plasmid vector, and characterized by ABI Big Dye terminator sequencing (Perkin Elmer; Foster City, CA). The nucleotide data reported will appear in DDBJ, EMBL, and GenBank nucleotide sequence databanks under the accession number AJ243821.
RNA Preparation and Reverse Transcriptase-assisted PCR
RNAs were prepared from mouse tissues by the CsCl method (
The oligonucleotide primers used for detection of Aire transcripts were CAGCAACTCTGGCCTCAAAG and CTTC-GAACTTGTTGGGTGTATAA (GenBank entry AJ243821; nucleotides 431450 and 693715, respectively).
mRNA In Situ Hybridization
Adult NMRI mice were anesthetized with pentobarbital and perfused transcardially with physiological saline, followed by 4% paraformaldehyde in PBS for 4 min. Then the tissues were excised and immersed in the same fixative for an additional 60 min. Thereafter, the tissues were processed for paraffin embedding and 5-µm sections were mounted on Polysine glasses (Menzel; Braunschweig, Germany). The sections were deparaffinized, rehydrated, dehydrated, and dried. The sections were hybridized in a hybridization cocktail containing a mixture of three different 33P-labeled (Life Science Products; Boston, MA) cRNA probes (1 x 107 cpm/ml) covering mouse Aire cDNA sequence nucleotides 1421, 542879, and 12871906 (accession number AJ243821) for 18 hr at 55C. Then the slides were washed twice (15 min) in 2 x SSC at room temperature, treated with 100 µg/ml RNase A (Ambion; Austin, TX) for 30 min at 37C, and washed with 0.5 x SSC and 0.1 x SSC at 60C for 15 min each. The slides were dipped in water, dehydrated through graded ethanol, and either covered with Kodak MR5 (Eastman Kodak; Rochester, NY) autoradiography film or dipped in Kodak NTB2 emulsion. The films were exposed for 260 days, developed with Kodak LX 24 developer, and fixed with AL4 (Eastman Kodak) fixative. The dipped sections were exposed for 60 days and develop with D19 (Eastman Kodak) developer, fixed with G333 (Agfa Gevaert; Cologne, Germany) fixative, counterstained with cresyl violet, and coverslipped. The mixture of the sense probes was used as a negative control. Additional control experiments were carried out by treatment of the tissue sections with RNase A (100 µg/ml) for 30 min at 37C before hybridization with antisense probes.
Cell Culture and Transfections
The pAP42 construct containing the full-length mouse Aire cDNA in pcDNA3.1+ (Invitrogen; Carlsbad, CA) was transfected into cultured cell lines by using the FuGene 6 transfection reagent according to the manufacturer's instructions (Roche Diagnostics; Indianapolis, IN). Altogether, five different cell lines (human 293, monkey CV-1 and COS-1, mouse NIH3T3, and hamster BHK cells) were used in this study. For indirect immunofluorescence detection, cells were fixed with 3.5% paraformaldehyde 48 hr after transfection. The fixed cells were permeabilized with 0.1% Triton X-100 and blocked with 0.2% BSA. Tetramethylrhodamine isothiocyanate (TRITC)-conjugated anti-rabbit IgG (Jackson ImmunoResearch Laboratories; West Grove, PA) were used as secondary antibodies.
Production of the Mouse APECED Protein-specific Polyclonal Antisera
The peptide sequence KTKPPKKPDGNLESQHL, corresponding to amino acids 160176 of mouse APECED protein, was chosen for production of antisera. Immunizations were performed with custom-synthesized MAP peptides (Research Genetics; Huntsville, AL). The immune sera were affinity-purified using respective synthetic peptides attached to CNBr-activated Sepharose 4B columns according to the manufacturer's instructions (Amersham Pharmacia Biotech; Piscataway, NJ).
Western Blotting
The tissue culture samples for SDS-PAGE electrophoresis were collected 48 hr after transfection. Proteins were separated on 11% SDS-PAGE and analyzed by Western blotting with specific polyclonal antisera raised against a synthetic peptide corresponding to amino acids 160176 of the mouse APECED protein.
Immunohistochemistry
For immunohistochemistry, adult mice were fixed as described for in situ hybridization. The samples were cryoprotected with 15% sucrose in PBS, frozen with carbon dioxide, and 10-µm sections were cut. Mouse peripheral blood leukocytes (PBLs) were separated by centrifugation, smeared onto Polysine glasses, and fixed by immersion in 4% paraformaldehyde for 5 min. Endogenous peroxidase activity was inhibited by immersing the sections in 0.3% hydrogen peroxide in PBS for 520 min. The sections were incubated with rabbit affinity-purified anti-peptide antibody raised against mouse APECED protein (amino acids 160176; dilution 1:100200) at 4C overnight in a buffer containing 1% BSA and 0.3% Triton X-100. Then the sections were incubated with biotinylated goat anti-rabbit IgG (diluted 1:300) and ABC complex (Vectastain Elite Kit; Vector Laboratories, Burlingame, CA) for 30 min each. The immunoreaction was visualized with nickel-intensified DAB as a chromogen. The sections were then dehydrated and embedded in Entellan. Controls included preimmune serum and antibody presaturated with 20 µg/ml of the peptide used for immunization. In addition, extracts prepared from COS-1 cells transfected with full-length Aire cDNA and empty expression vector were used for preadsorbtion experiments.
In Vitro Translation
The full-length Aire cDNA in Bluescript SK+ was in vitro translated using the TNT-coupled Reticulocyte Lysate System according to the manufacturer's instructions (Promega; Madison, WI). The protein samples were analyzed on an 11% SDS-PAGE, followed by autoradiography.
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Results |
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Mouse Aire cDNA Cloning
The mouse cDNA library was screened with human AIRE cDNA as a probe. Positive clones were plaque-purified through subsequent screening cycles and characterized by nucleotide sequencing. One of the isolated clones encoded for amino acids 430552 of mouse APECED protein, as deduced from the mouse Aire genomic DNA sequence (
Expression of Aire cDNA in Mammalian Tissue Culture Cells
The subcellular localization of mouse APECED protein was studied by transient expression and indirect immunofluorescence experiments using the antiserum raised against a synthetic APECED peptide. The analysis of different cell types (BHK, NIH3T3, CV-1, COS-1, and 293) revealed three different staining patterns: (a) speckled nuclear distribution excluding the nucleoli (Fig 1A); (b) filamentous cytoplasmic pattern resembling the staining characteristic to vimentin (Fig 1B); and (c) both of these two distributions in the same transfected cell (Fig 1C). In BHK cells, over 30% of the transfected cells had a speckled nuclear APECED protein distribution, 30% exhibited filamentous cytoplasmic staining, 30% exhibited both distributions, and the remaining cells had other staining patterns, such as clustering of the protein in the nuclear envelope. In mouse NIH3T3 cells, over 70% of the transfected cells showed cytoplasmic staining only. Speckled nuclear staining was observed in less than 10% of transfected cells. A mixed localization pattern was observed in less than 10% of the cells, and the rest exhibited other staining patterns. In addition, the CV-1, COS-1, and 293 cell lines transiently transfected with mouse Aire cDNA displayed the different staining patterns mentioned above (data not shown). The staining pattern varied among the different cell lines. The CV-1 and COS-1 cells had a predominant filamentous cytoplasmic staining pattern, whereas in the majority of transfected 293 cells APECED protein was localized in nuclear speckles. The subcellular distribution of mouse APECED protein therefore closely resembles that of its human counterpart (
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Analysis of Aire mRNA Expression by RT-PCR and mRNA In Situ Hybridization
The expression of mouse Aire was first studied by RT-PCR. Analysis of mouse total RNA revealed the amplification of Aire-specific transcripts in all the tissues studied, i.e., thymus, spleen, lymph node, liver, kidney, testis, brain, and fetal liver (data not shown). In general, our RT-PCR results are in agreement with recently published studies (
Aire expression in mouse tissues was further studied by mRNA in situ hybridization. Our initial experiments with radioactively labeled oligonucleotides failed to detect Aire transcripts even with rather long exposure times (data not shown). To improve the sensitivity of our assays, we decided to use radioactively labeled mouse Aire cRNA as the hybridization probe. Additional improvement in sensitivity was achieved by using a mixture of three cRNA probes covering different parts of the Aire transcript (see Materials and Methods). In the thymus, a strong signal for Aire mRNA could be seen in a small population of cells in the medulla. These cells were larger than thymocytes, stained lightly with cresyl violet, and obviously represented reticular (epithelial) cells of the thymus. Part of the epithelial cells clearly lacked Aire mRNA. A moderate level of staining was also seen in a small population of medullary thymocytes (Fig 2A2E).
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Our results on Aire transcript expression in mouse thymus are in agreement with recently published results showing scattered cells expressing the AIRE mRNA in the medullary zone of the thymus of 17-day mouse embryo and in human juvenile thymus (
Immunohistochemistry
To gain information on the distribution of the APECED protein at the cellular level, we carried out immunohistochemical stainings of mouse tissue sections with antisera raised against a synthetic polypeptide corresponding to mouse APECED protein. The specificity of the immunohistochemical reaction was demonstrated by the lack of immunostaining of tissue sections incubated with antisera preadsorbed either with the synthetic peptide used for immunization (Fig 5M, Fig 5O, and Fig 5P) or extracts prepared from COS-1 cells transfected with full-length Aire cDNA (Fig 5N). The staining was not affected in control experiments performed with antisera preadsorbed with the extracts prepared from COS-1 cells transfected with empty expression vector (data not shown). The specificity of the antiserum was further studied by staining the Western blots of the lysates of NIH-3T3 and COS-1 cell lines transiently transfected with Aire cDNA (Fig 3). A single major protein of the expected molecular weight can be seen in Aire-transfected cell lines (Fig 3, Lane A), whereas cells transfected with empty expression vector remain completely unstained (Fig 3, Lane C). Additional evidence for the specificity of the antiserum was obtained by staining Aire-knockout mouse tissues. The thymus, liver, and cerebellum of the Aire-knockout mouse stained with Aire-specific antiserum did not show any staining (data not shown; Puhakka et al. unpublished observations).
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Immune System. A small number of intensely Aire-immunoreactive (Aire-IR) cells was observed in the medulla of the thymus. These cells were larger than thymocytes and had large non-labeled cytoplasm characteristic of reticular (epithelial) cells. The labeling was localized in distinct dots in the nucleus (Fig 4A). Some of the reticular cells were devoid of labeling. In addition, the cells in the thymic corpuscles were intensely stained. A small number of medullary thymocytes exhibited low to moderate staining (Fig 4B). Cortical thymocytes were devoid of immunostaining.
In the red pulp of spleen, large cells possessing prominent nuclei were clearly labeled. These cells most probably represent tissue macrophages (Fig 4C). In addition, a large number of lymphocytes and some neutrophilic granulocytes in the red pulp were Aire-IR (Fig 4C). A few labeled reticular cells could be observed. Light staining was seen in the smooth muscle cells of the trabecules. The white pulp of spleen was devoid of immunostaining.
In the medulla of the lymph nodes, a population of the lymphocytes was clearly stained, but the cortical germinal centers remained non-stained. Some labeled reticular cells could be seen in the medulla. Mast cell nuclei appeared to be unlabeled, although the cytoplasmic granules exhibited strong nonspecific staining which sometimes obscured the nucleus (Fig 4D).
In bone marrow, a subpopulation of large megacaryocytes showed intense staining. The labeling was localized to distinct dots in the nuclei. In addition, scattered smaller cells exhibited clear staining; these cells probably represent lymphoblasts and myeloblasts (Fig 4E).
In the peripheral blood smears, strong nuclear staining could be seen in lymphocytes, polymorphonuclear leukocytes, and monocytes. A small population of lymphocytes appeared to be devoid of staining (Fig 4F).
Urinary Tract. In kidney, the epithelial cells of the proximal and distal convoluted tubules were labeled. The loops of Henle and collecting ducts were mainly unlabeled. The dark cells of the collecting tubules exhibited cytoplasmic staining close to the cell membrane. The podocytes of glomeruli were slightly labeled. Weak staining was also seen in the transitional epithelium in the kidney pelvis (Fig 4G).
In urinary bladder, the transitional epithelium was strongly Aire-IR. The smooth muscle cells of bladder exhibited weak staining (Fig 4H).
Genital Organs. In the testis, immunostaining could be seen in a stage-specific manner in a few seminiferous tubules. Staining was localized in distinct dots in the nuclei of pachytene spermatocytes and round spermatids. In a few tubules, some spermatogonia and Sertoli cells showed Aire-IR (Fig 4I). In the interstitial tissue, most of the cells exhibited nuclear staining. A population of these cells also showed moderate cytoplasmic staining, which was abolished by presaturation (Fig 4J). In epididymis, seminal vesicle (Fig 4K), and prostate the nuclei of epithelial cells stained strongly.
In the ovary, nuclear staining could be observed in the granular cells of different size follicles. The oocytes were also labeled. The cells of corpora lutea and interstitial tissue exhibited strong cytoplasmic staining without clear nuclear labeling. This staining could not be abolished by presaturation of the antibody (Fig 4L). In the uterus, the epithelial cells of the mucosa and secretory glands were clearly stained. The smooth muscle cells of the myometrium were also immunopositive.
Endocrine Organs. Large numbers of cells in anterior and intermediate lobes of pituitary were Aire-IR (Fig 4M). In the adrenal gland, clear nuclear staining could be observed in the cells of zona glomerulosa and in the medullar chromaffin cells (Fig 4N). The cells of the zona fasciculata and reticulata exhibited strong cytoplasmic staining without a clear nuclear signal. This cytoplasmic staining was abolished by presaturation (data not shown). Most of the cells in the adrenal medulla displayed clear staining (Fig 4O). In the thyroid gland, a variable degree of immunostaining could be seen in the epithelial cells of the follicles. Clear staining was observed in parafollicular cells (Fig 4P).
Alimentary Tract. In the salivary glands, most of the secretory cells of the acini and secretory ducts were moderately to strongly Aire-IR (Fig 5A). In the liver, most of the hepatocytes were very intensely Aire-IR, although some were devoid of labeling (Fig 5B). The staining intensity was similar in different regions of the liver lobule. Küppfer cells were also stained (Fig 5B). The staining pattern of the hepatocyte nuclei was uneven and resembled that of transiently transfected cell lines (Fig 5C). The epithelial cells and glandular cells in different parts of the alimentary canal were Aire-IR (Fig 5D). In the pancreas, both the exocrine and endocrine cells were Aire-IR (Fig 5E).
Respiratory System. In the respiratory tract, Aire immunoreactivity could be seen in the respiratory epithelium of large airways from the larynx to the respiratory bronchioles (Fig 5F and Fig 5H). The alveolar Type I and Type II cells displayed weak staining, and alveolar macrophages were clearly labeled. In tracheal cartilage, the undifferentiated cells in the perichondrium and differentiating chondroblasts were stained, but mature chondrocytes were not labeled (Fig 5G).
Nervous System. APECED immunostaining was widely distributed in the central nervous system. The cerebral cortex, hippocampus (Fig 5I), amygdala, thalamic nuclei, hypothalamus, cerebellar cortex, various brainstem nuclei, spinal cord, and dorsal root ganglia (Fig 5K) all revealed strong immunoreactivity. In the cerebellum, intense Aire-IR was observed in the nuclei of the Purkinje cells, while the granular neurons exhibited modest staining (Fig 5J). The most prominent staining was in neurons, but different types of glial cells were also labeled. In the eye, the ganglion cells and bipolar neurons of the retina showed moderate staining, but the rods and cones were negative (Fig 5L).
Those tissues in which Aire expression was detected with at least two different experimental techniques are listed in Table 1.
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Discussion |
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To understand the biological function of APECED protein, we studied the spatial expression pattern of the Aire gene in mouse using several different techniques such as RT-PCR, in situ mRNA hybridization, and immunohistochemical staining. Our results revealed expression of Aire in different cell populations in a broad range of rather different types of mouse tissues. Because APECED is an autoimmune disorder, the immunologically relevant organs would be expected to express Aire. In the thymus, spleen, lymph nodes, and bone marrow, Aire expression was detected in different cell populations of lymphoid and myeloid lineages. It should be noted that no double stainings with specific cell-surface markers were performed and therefore the precise identity and stage of differentiation of cells expressing Aire remain to be determined. In addition the reticular epithelial cells and corpuscle cells of the thymus and epithelial cells of the lymph vessels expressed Aire. Our findings in thymus, spleen, and lymph nodes are in a good agreement with earlier studies on human AIRE expression by us and others (
Outside of the immune system, the APECED protein was detected in the brain, liver, kidney, pancreas, intestinal canal, gonads, pituitary, thyroid, and adrenal glands. The cells expressing the APECED protein represent rather diverse cell types, ranging from epithelial cells in several different organs to neurons and glial cells in the central nervous system. Our results showing a broad range of tissues expressing APECED protein are further supported by our studies of the rat. Immunohistochemical staining of rat tissues with antiserum raised against human APECED protein have revealed an almost identical expression pattern as that observed in the mouse (unpublished results).
In all the cell types that showed nuclear labeling in adult mouse tissues, the staining was unevenly distributed and resembled the speckled, nuclear body-like staining that we observed in mammalian cell cultures transfected with mouse Aire cDNA. Similar results have been obtained in experiments with the human counterpart of Aire (
It should be noted that in a few cell types, such as interstitial cells of the testis and few neurons in the trigeminal ganglion, cytoplasmic staining was observed in addition to nuclear labeling. Our results show that, in transiently transfected tissue culture cells, mouse APECED protein also has a cytoplasmic vimentin-like distribution. The proportion of the different distribution types varies depending on the cell line used in the transient expression. It has been speculated by
It has been suggested that APECED protein might be involved in the regulation of self-tolerance either by clonal deletion in the thymus or by induction of peripheral T-cell anergy (
Conclusions
In this study, we have shown by in situ hybridization and immunohistochemistry that Aire is widely expressed throughout the entire organism. For the first time, we have shown the presence of the APECED protein in multiple tissues outside the immune system in very diverse cell types. We have shown that APECED protein has dual subcellular localization in transiently transfected cell lines. In vivo it was predominantly located in nuclei. However, in a few cell types cytoplasmic localization was also observed. Our results imply that APECED protein most likely has different biological functions in different tissues depending on the tissue- and cell type-specific background. Identification of the protein partners interacting with APECED protein and the cellular pathways regulated by its activity will be of critical importance to understanding the biological role of the APECED protein. It will also provide us with valuable information on the molecular mechanisms behind APECED pathology and new insights into autoimmune diseases in general.
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Footnotes |
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1 Present address: Gonda Neuroscience and Genetics Research Center, UCLA Department of Human Genetics, Los Angeles, CA.
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Acknowledgments |
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Supported by the Academy of Finland, the Ulla Hjelt fond of the Foundation for Pediatric Research, the Medical Research Fund of Tampere University Hospital, and by resources from the Helsinki Biomedical Graduate School MD/PhD program.
The excellent technical assistance of Ulla-Margit Jukarainen, Tuula Airaksinen, Anne Vikman, and Katri Miettinen is greatly appreciated. Donald Smart is acknowledged for language revision.
Received for publication March 28, 2000; accepted September 13, 2000.
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Literature Cited |
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![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Aasland R, Gibson TJ, Stewart AF (1995) The PHD finger: implications for chromatin-mediated transcriptional regulation. Trends Biochem Sci 20:56-59[Medline]
Björses P, Halonen M, Palvimo JJ, Kolmer M, Aaltonen J, Ellonen P, Perheentupa J, Ulmanen I, Peltonen L (2000) Mutations in the AIRE gene: effects on sub-cellular location and transactivation function of the APECED protein. Am J Hum Genet 66:378-392[Medline]
Björses P, Pelto-Huikko M, Kaukonen J, Aaltonen J, Peltonen L, Ulmanen I (1999) Localization of the APECED protein in distinct nuclear structures. Hum Mol Genet 8:259-266
Blechschmidt K, Schweiger M, Wertz K, Poulson R, Christensen HM, Rosenthal A, Lehrach H, Yaspo ML (1999) The mouse aire gene: comparative genomic sequencing, gene organization, and expression. Genome Res 9:158-166
Bloch DB, Chiche JD, Orth D, de la Monte SM, Rosenzweig A, Bloch KD (1999) Structural and functional heterogeneity of nuclear bodies. Mol Cell Biol 19:4423-4430
Chirgwin J, Przybyla AE, McDonald R, Rutter W (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294-5299[Medline]
An autoimmune disease, APECED, caused by mutations in a novel gene featuring two PHD-type zinc-finger domains. The Finnish-German APECED Consortium. Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy. (1997) Nature Genet 17:399-403[Medline]
Ganfornina MD, Sanchez D (1999) Generation of evolutionary novelty by functional shift. Bioessays 21:432-439[Medline]
Gibson TJ, Ramu C, Gemund C, Aasland R (1998) The APECED polyglandular autoimmune syndrome protein, AIRE-1, contains the SAND domain and is probably a transcription factor. Trends Biochem Sci 23:242-244[Medline]
Heery DM, Kalkhoven E, Hoare S, Parker MG (1997) A signature motif in transcriptional co-activators mediates binding to nuclear receptors. Nature 387:733-736[Medline]
Heino M, Peterson P, Kudoh J, Nagamine K, Lagerstedt A, Ovod V, Ranki A, Rantala I, Nieminen M, Tuukkanen J, Scott HS, Antonarakis SE, Shimizu N, Krohn K (1999a) Autoimmune regulator is expressed in the cells regulating immune tolerance in thymus medulla. Biochem Biophys Res Commun 257:821-825[Medline]
Heino M, Scott HS, Chen Q, Peterson P, Mäenpää U, Papasavvas MP, Mittaz L, Barras C, Rossier C, Chrousos GP, Stratakis CA, Nagamine K, Kudoh J, Shimizu N, Maclaren N, Antonarakis SE, Krohn K (1999b) Mutation analyses of North American APS-1 patients. Hum Mutat 13:69-74[Medline]
Kozak M (1991) An analysis of vertebrate mRNA sequences: intimations of translational control. J Cell Biol 115:887-903[Abstract]
Matera AG (1999) Nuclear bodies: multifaceted subdomains of the interchromatin space. Trends Cell Biol 9:302-309[Medline]
Mittaz L, Rossier C, Heino M, Peterson P, Krohn KJ, Gos A, Morris MA, Kudoh J, Shimizu N, Antonarakis SE, Scott HS (1999) Isolation and characterization of the mouse aire gene. Biochem Biophys Res Commun 255:483-490[Medline]
Mu ZM, Chin KV, Liu JH, Lozano G, Chang KS (1994) PML, a growth suppressor disrupted in acute promyelocytic leukemia. Mol Cell Biol 14:6858-6867[Abstract]
Nagamine K, Peterson P, Scott HS, Kudoh J, Minoshima S, Heino M, Krohn KJ, Lalioti MD, Mullis PE, Antonarakis SE, Kawasaki K, Asakawa S, Ito F, Shimizu N (1997) Positional cloning of the APECED gene. Nature Genet 17:393-398[Medline]
Pearce SH, Cheetham T, Imrie H, Vaidya B, Barnes ND, Bilous RW, Carr D, Meeran K, Shaw NJ, Smith CS, Toft AD, Williams G, KendallTaylor P (1998) A common and recurrent 13-bp deletion in the autoimmune regulator gene in British kindreds with autoimmune polyendocrinopathy type 1. Am J Hum Genet 63:1675-1684[Medline]
Pearson M, Carbone R, Sebastiani C, Cioce M, Fagioli M, Saito S, Higashimoto Y, Appella E, Minucci S, Pandolfi PP, Pelicci PG (2000) PML regulates p53 acetylation and premature senescence induced by oncogenic Ras. Nature 406:207-210[Medline]
Peterson P, Nagamine K, Scott H, Heino M, Kudoh J, Shimizu N, Antonarakis SE, Krohn KJ (1998) APECED: a monogenic autoimmune disease providing new clues to self-tolerance. Immunol Today 19:384-386[Medline]
Pitkänen J, Doucas V, Sternsdorf T, Nakajima T, Aratani S, Jensen K, Will H, Vähämurto P, Ollila J, Vihinen M, Scott HS, Antonarakis SE, Kudoh J, Shimizu N, Krohn K, Peterson P (2000) The autoimmune regulator protein has transcriptional transactivating properties and interacts with the common coactivator CREB-binding protein. J Biol Chem 275:16802-16809
Rinderle C, Christensen HM, Schweiger S, Lehrach H, Yaspo ML (1999) AIRE encodes a nuclear protein co-localizing with cytoskeletal filaments: altered sub-cellular distribution of mutants lacking the PHD zinc fingers. Hum Mol Genet 8:277-290
Rosatelli MC, Meloni A, Meloni A, Devoto M, Cao A, Scott HS, Peterson P, Heino M, Krohn KJ, Nagamine K, Kudoh J, Shimizu N, Antonarakis SE (1998) A common mutation in Sardinian autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy patients. Hum Genet 103:428-434[Medline]
Ruan QG, Wang CY, Shi JD, She JX (1999) Expression and alternative splicing of the mouse autoimmune regulator gene (Aire). J Autoimmun 13:307-313[Medline]
Scott HS, Heino M, Peterson P, Mittaz L, Lalioti MD, Betterle C, Cohen A, Seri M, Lerone M, Romeo G, Collin P, Salo M, Metcalfe R, Weetman A, Papasavvas MP, Rossier C, Nagamine K, Kudoh J, Shimizu N, Krohn KJ, Antonarakis SE (1998) Common mutations in autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy patients of different origins. Mol Endocrinol 12:1112-1119
Sternsdorf T, Grotzinger T, Jensen K, Will H (1997a) Nuclear dots: actors on many stages. Immunobiology 198:307-331[Medline]
Sternsdorf T, Jensen K, Reich B, Will H (1999) The nuclear dot protein sp100, characterization of domains necessary for dimerization, subcellular localization, and modification by small ubiquitin-like modifiers. J Biol Chem 274:12555-12566
Sternsdorf T, Jensen K, Will H (1997b) Evidence for covalent modification of the nuclear dot-associated proteins PML and Sp100 by PIC1/SUMO-1. J Cell Biol 139:1621-1634
Torii S, Egan DA, Evans RA, Reed JC (1999) Human Daxx regulates Fas-induced apoptosis from nuclear PML oncogenic domains (PODs). EMBO J 18:6037-6049
Wang CY, DavoodiSemiromi A, Huang W, Connor E, Shi JD, She JX (1998) Characterization of mutations in patients with autoimmune polyglandular syndrome type 1 (APS1). Hum Genet 103:681-685[Medline]
Wang CY, Shi JD, DavoodiSemiromi A, She JX (1999) Cloning of aire, the mouse homologue of the autoimmune regulator (AIRE) gene responsible for autoimmune polyglandular syndrome type 1 (APS1). Genomics 55:322-326[Medline]
Ward L, Paquette J, Seidman E, Huot C, Alvarez F, Crock P, Delvin E, Kampe O, Deal C (1999) Severe autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy in an adolescent girl with a novel AIRE mutation: response to immunosuppressive therapy. J Clin Endocrinol Metab 84:844-852
Zheng P, Guo Y, Niu Q, Levy DE, Dyck JA, Lu S, Sheiman LA, Liu Y (1998) Proto-oncogene PML controls genes devoted to MHC class I antigen presentation. Nature 396:373-376[Medline]