Alopecia universalis associated with a mutation in the human hairless gene

Science

Volumn 279, Issue 5351, 1998, Pages 720-724

  Ahmad, Wasim ^a   Ul Haque, Muhammad Faiyaz ^b   Brancolini, Valeria ^c   Tsou, Hui C ^a   Ul Haque, Sayed ^b   Lam, Hamut ^a   Aita, Vincent M ^a   Owen, Jason ^d   DeBlaquiere, Michelle ^d   Frank, Jorge ^a   Cserhalmi Friedman, Peter B ^a   Leask, Andrew ^e   McGrath, John A ^f   Peacocke, Monica ^a   Ahmad, Mahmud ^b   Ott, Jurg ^c   Christiano, Angela M ^a  

^a Och Spine at New York Presbyterian Hospitals   (United States)

^b QUAID I AZAM UNIVERSITY   (Pakistan)

^c ROCKEFELLER UNIVERSITY   (United States)

^d Research Genetics   (United States)

^e FIBROGEN INC   (United States)

^f ST THOMAS HOSPITAL   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

TRANSCRIPTION FACTOR; ZINC FINGER PROTEIN;

ALOPECIA; ARTICLE; CHROMOSOME 8P; GENETIC LINKAGE; HUMAN; MISSENSE MUTATION; PRIORITY JOURNAL;

ALOPECIA; AMINO ACID SEQUENCE; ANIMALS; BRAIN; CHROMOSOME MAPPING; CHROMOSOMES, HUMAN, PAIR 8; DNA-BINDING PROTEINS; FEMALE; FORKHEAD TRANSCRIPTION FACTORS; GENE EXPRESSION; GENES, RECESSIVE; HOMOZYGOTE; HUMANS; MALE; MICE; MICE, INBRED HRS; MICROSATELLITE REPEATS; MOLECULAR SEQUENCE DATA; MUTATION; PEDIGREE; PROTEINS; RATS; SEQUENCE ANALYSIS, DNA; SEQUENCE HOMOLOGY, AMINO ACID; SKIN; TRANSCRIPTION FACTORS; ZINC FINGERS;

MURINAE;

EID: 6844265562     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.279.5351.720     Document Type: Article

References (37)

1. M. H. Hardy, Trends Genet. 8, 159 (1992); T. A. Rosenquist and G. R. Martin, Dev. Dyn. 205, 379 (1996).
2. M. H. Hardy, Trends Genet. 8, 159 (1992); T. A. Rosenquist and G. R. Martin, Dev. Dyn. 205, 379 (1996).
3. J. M. Hebert, T. Rosenquist, J. Gotz, G. R. Martin, Cell 78, 1017 (1994).
4. L. Guo, L. Degenstein, E. Fuchs, Genes Dev. 10, 165 (1996).
5. P. Zhou, C. Byrne, J. Jacobs, E. Fuchs, ibid. 9, 700 (1995).
6. P. J. Koch et al., J. Cell Biol. 137, 1091 (1997).
7. M. Nehls, D. Pfeifer, M. Schorpp, H. Hedrich, T. Boehm, Nature 372, 103 (1994); J. Segre, J. L. Nemhauser, B. A. Taylor, J. H. Nadeau, E. S. Lander, Genomics 28, 549 (1995); K. Schüddenkopf, M. Schorpp, T. Boehm, Proc. Natl. Acad. Sci. U.S.A. 93, 9661 (1996).
8. M. Nehls, D. Pfeifer, M. Schorpp, H. Hedrich, T. Boehm, Nature 372, 103 (1994); J. Segre, J. L. Nemhauser, B. A. Taylor, J. H. Nadeau, E. S. Lander, Genomics 28, 549 (1995); K. Schüddenkopf, M. Schorpp, T. Boehm, Proc. Natl. Acad. Sci. U.S.A. 93, 9661 (1996).
9. M. Nehls, D. Pfeifer, M. Schorpp, H. Hedrich, T. Boehm, Nature 372, 103 (1994); J. Segre, J. L. Nemhauser, B. A. Taylor, J. H. Nadeau, E. S. Lander, Genomics 28, 549 (1995); K. Schüddenkopf, M. Schorpp, T. Boehm, Proc. Natl. Acad. Sci. U.S.A. 93, 9661 (1996).
10. A. Rook and K. Dawber, Diseases of the Hair and Scalp (Blackwell, Oxford, UK, ed. 2, 1991), p.136; W. F. Bergfeld, Am. J. Med. 98, 95S (1995).
11. A. Rook and K. Dawber, Diseases of the Hair and Scalp (Blackwell, Oxford, UK, ed. 2, 1991), p.136; W. F. Bergfeld, Am. J. Med. 98, 95S (1995).
12. H. K. Muller et al., Br. J. Dermatol. 102, 609 (1980).
13. V. C. Sheffield, D. Y. Nishimura, E. M. Stone, Curr. Opin. Genet. Dev. 5, 335 (1995).
14. Blood samples were collected from 36 members of the AU family, according to local informed consent procedures. DNA was isolated according to standard techniques (29). Fluorescent automated genotyping for the genome-wide linkage search was carried out at as a service by Research Genetics, with 386 markers covering the genome at about 10-cM intervals.
15. 33P]deoxyadenosine triphosphate; a PCR reaction consisting of 7 min at 95°C, followed by 27 cycles of 1 min at 95°C, 1 min at 55°C, and 1 min at 72°C; and electrophoresis in a 6% polyacrylamide gel (Sequa-gel; Action Scientific, Atlanta, GA). Microsatellite markers were visualized by exposure of the gel to autoradiography, and genotypes were assigned by visual inspection.
16. Statistical calculations for linkage analysis were carried out with the computer program FASTLINK version 3.OP [A. A. Schaffer, Hum. Hered. 46, 226 (1996)], which enables all inbreeding loops in the family to be retained and has the capability for twopoint analysis. Autosomal recessive with complete penetrance was assumed with a disease allele frequency of 0.0001. The lod scores were calculated with equal allele frequencies; however, results did not change when the frequency of the marker allele in association with the disease allele was set as high as 0.9. Multipoint analysis was not possible because of the large number of inbreeding loops and the complexity of the pedigree.
17. H. C. Brooke, J. Hered. 15, 173 (1924); M. B. Cachon-Gonzalez et al., Proc. Natl. Acad. Sci. U.S.A. 91, 7717 (1994).
18. H. C. Brooke, J. Hered. 15, 173 (1924); M. B. Cachon-Gonzalez et al., Proc. Natl. Acad. Sci. U.S.A. 91, 7717 (1994).
19. For RT-PCR of human hairless cDNA sequences, total RNA was extracted from cultured skin fibroblasts from hair-bearing skin from a control individual according to standard methods (29). We reverse transcribed human hairless mRNAs with mouse mammary leukemia virus RT (Gibco-BRL), using an oligo-deoxyribosylthymine primer (Pharmacia). PCR was carried out with the following primers, constructed on the basis of the mouse hairless sequence (GenBank accession number Z32675): 5′-TGAGGGCTCTGTCCTCCTGC-3′ (sense) and 5′-GCTGGCTCCCTGGTGGTAGA-3′ (antisense). PCR conditions were 5 min at 95°C, followed by 35 cycles of 1 min at 95°C, 1 min at 55°C, and 1 min at 72°C, with AmpliTaq Gold DNA polymerase (Perkin-Elmer). After direct sequencing of the human cDNA, exonbased primers were designed and used to amplify genomic DNA as template and directly sequenced with the ABI 310 Automated Sequencer. The intronexon borders were determined by comparison of the cDNA and the genomic DNA sequences at both the 5′ donor and 3′ acceptor splice junctions. The human hairless sequence has been deposited in GenBank (accession number AFO39196).
20. C. C. Thompson, J. Neurosci. 16, 7832 (1996).
21. + mRNA was hybridized with the same probe under identical conditions.
22. See the Genome Data Base at www.bis.med. jhmi.edu
23. A segment of human hairless intron 13 was PCR-amplified and used for radiation hybrid mapping with the G3 panel by Research Genetics. Primers were as follows: 5'-TATGTCACCAAGGGCCAGCC-3′ (sense) and 5′-TCAGGGTAGGGGGTCATGCC-3′ (antisense). PCR conditions were 5 min at 95°C, followed by 35 cycles of 1 min at 95°C, 1 min at 55°C, and 1 min at 72°C, with AmpliTaq Gold DNA polymerase (Perkin-Elmer). PCR primers specifically amplified human hairless and did not crosshybridize with the hamster DNA used in the radiation hybrid panel.
24. See www.marshmed.org/genetics/maps/ss-maps/ 8ss.txt
25. See www.shgc.stanford.edu
26. Primers for specific amplification of exon 15 were placed in the flanking introns: 5′-AGTGCCAGGATTACAGGCGT-3′ (sense, intron 15) and 5′-CTGAGGAGGAAAGAGCGCTC-3′ (sense, intron 16). PCR fragments were purified on AGTC Centriflex columns (Edge BioSystems, Gaithersburg, MD) and sequenced directly with POP-6 polymer on an ABI Prism 310 Automated Sequencer (Perkin-Elmer). The mutation was verified by restriction endonuclease digestion with Hga I, according to the manufacturer's specifications (New England Biolabs).
27. A. Ganguly, M. J. Rock, D. J. Prockop, Proc. Natl. Acad. Sci. U.S.A. 90, 10325 (1993).
28. J. P. Stoye, S. Fenner, G. E. Greenoak, J. M. Moran, Cell 54, 383 (1988).
29. R. J. Arced, A. A. J. King, M. C. Simon, S. H. Orkin, D. B. Wilson, Mol. Cell. Biol. 13, 2235 (1993).
30. C. Höög, M. Schalling, E. Grunder-Brundell, B. Daneholt, Mol. Reprod. Dev. 30, 173 (1991).
31. P. J. Morrissey, D. R. Parkinson, R. S. Schwartz, S. D. Waksal, J. Immunol. 125, 1558 (1980).
32. C. H. Gallagher, F. R. C. Path, P. J. Canfield, G. E. Greenoak, V. E. Reeve, J. Invest. Dermatol. 83, 169 (1984).
33. G. L. Semenza, Hum. Mutat. 3, 180 (1994); D. S. Latchman, N, Engl. J. Med. 334, 28 (1996); D. Engelkamp and V. van Heyningen, Curr. Opin. Genet. Dev. 6, 334 (1996).
34. G. L. Semenza, Hum. Mutat. 3, 180 (1994); D. S. Latchman, N, Engl. J. Med. 334, 28 (1996); D. Engelkamp and V. van Heyningen, Curr. Opin. Genet. Dev. 6, 334 (1996).
35. G. L. Semenza, Hum. Mutat. 3, 180 (1994); D. S. Latchman, N, Engl. J. Med. 334, 28 (1996); D. Engelkamp and V. van Heyningen, Curr. Opin. Genet. Dev. 6, 334 (1996).
36. J. Sambrook, E. G. Fritsch, T. Maniatis, Molecular Cloning, A Laboratory Manual, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, ed. 2, 1989).
37. We sincerely thank the family members for their participation in this work, S. Malik and S. M. Zaidi for their assistance during our stay in Chakwal, B. J. Longley for expert dermatopathological advice, and M. Grossman and P. Schneiderman for their infinite patience. Informed consent for the publication of the photographs as well as the pedigree was obtained through personal visits to the family members. Supported in part by grants from Quaid-i-Azam University, Islamabad, Pakistan (M.A.), the National Alopecia Areata Foundation (A.M.C.), NIH-National Human Genome Research Institute HG-00008 (J.O.), and NIH-National Institute of Arthritis and Musculoskeletal and Skin Diseases Research Center P30AR44535 (M.P., J.O., and A.M.C.).