Document Detail


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.
MedLine Citation:
PMID:  2308937     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
T Mimori; Y Ohosone; N Hama; A Suwa; M Akizuki; M Homma; A J Griffith; J A Hardin
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
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:
Created Date:  1990-04-06     Completed Date:  1990-04-06     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1777-81     Citation Subset:  IM    
Affiliation:
Department of Medicine, Keio University School of Medicine, Tokyo, Japan.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/M30938
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MeSH Terms
Descriptor/Qualifier:
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:
AM07107/AM/NIADDK NIH HHS; AR32549/AR/NIAMS NIH HHS
Chemical
Reg. No./Substance:
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

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