| Isolation and characterization of cDNA encoding the 80-kDa subunit protein of the human autoantigen Ku (p70/p80) recognized by autoantibodies from patients with scleroderma-polymyositis overlap syndrome. | |
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MedLine Citation:
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PMID: 2308937 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Anti-Ku (p70/p80) autoantibodies in patients with scleroderma-polymyositis overlap syndrome recognize a 70-kDa/80-kDa protein heterodimer which binds to terminal regions of double-stranded DNA. In the present study, we isolated full-length cDNAs that encode the 80-kDa Ku subunit. Initial screening of a human spleen cDNA library with anti-Ku antibodies yielded a cDNA of 1.0 kilobase (kb) (termed K71) encoding a portion of the 80-kDa Ku polypeptide (identification based on immunological criteria). In RNA blots, this cDNA hybridized with two mRNAs of 3.4 and 2.6 kb. In rescreening of a cDNA library constructed from simian virus 40-transformed human fibroblast mRNA with the K71 cDNA as a hybridization probe, three positive clones were isolated, and that bearing the longest insert (termed Ku80-6) was selected for further characterization. In vitro transcription and translation experiments produced an immunoprecipitable polypeptide which comigrated with the 80-kDa Ku subunit. The Ku80-6 cDNA proved to be 3304 nucleotides in length, with an additional poly(A) tail, closely approximating the size of the larger mRNA. It contains a single long open reading frame encoding 732 amino acids (Mr = 82,713). The putative polypeptide has a high content of acidic amino acids and a region with periodic repeat of leucine in every seventh position which may form the "leucine zipper" structure. In genomic DNA blots, probes derived from the opposite ends of cDNA Ku80-6 hybridized with several nonoverlapping restriction fragments from human leukocyte DNA, indicating that the gene encoding the 80-kDa Ku polypeptide is divided into several exons by intervening sequences. |
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Authors:
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T Mimori; Y Ohosone; N Hama; A Suwa; M Akizuki; M Homma; A J Griffith; J A Hardin |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 87 ISSN: 0027-8424 ISO Abbreviation: Proc. Natl. Acad. Sci. U.S.A. Publication Date: 1990 Mar |
Date Detail:
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Created Date: 1990-04-06 Completed Date: 1990-04-06 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 7505876 Medline TA: Proc Natl Acad Sci U S A Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 1777-81 Citation Subset: IM |
Affiliation:
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Department of Medicine, Keio University School of Medicine, Tokyo, Japan. |
| Data Bank Information | |
Bank Name/Acc. No.:
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GENBANK/M30938 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Antigens, Nuclear* Antigens, Surface / genetics*, immunology Autoantibodies / immunology*, isolation & purification Autoimmune Diseases / genetics* Base Sequence Cloning, Molecular DNA / genetics*, isolation & purification DNA Helicases* DNA-Binding Proteins / genetics*, immunology Gene Library Humans Macromolecular Substances Molecular Sequence Data Molecular Weight Myositis / genetics*, immunology RNA, Messenger / genetics Restriction Mapping Scleroderma, Systemic / genetics*, immunology Spleen / immunology Syndrome |
| Grant Support | |
ID/Acronym/Agency:
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AM07107/AM/NIADDK NIH HHS; AR32549/AR/NIAMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Antigens, Nuclear; 0/Antigens, Surface; 0/Autoantibodies; 0/DNA-Binding Proteins; 0/Ku autoantigen; 0/Macromolecular Substances; 0/RNA, Messenger; 0/XRCC5 protein, human; 9007-49-2/DNA; EC 3.6.1.-/DNA Helicases |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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