Mutations in hamster single-strand break repair gene XRCC1 causing defective DNA repair

M. Richard Shen, Malgorzata Z. Zdzienicka, Harvey Mohrenweiser, Larry H. Thompson, Michael P. Thelen

Research output: Contribution to journalArticle

109 Citations (Scopus)

Abstract

The molecular basis for the DNA repair dysfunction observed in mutant Chinese hamster ovary cell lines of X-ray repair cross complementing group 1 (XRCC1) is unknown and the exact role of the XRCC1 protein remains unclear. To help clarify the role of the XRCC1 gene we analyzed four mutant cell lines of this complementation group and a revertant cell line for XRCC1 protein content and for sequence alterations in the XRCC1 coding region. Immunoblot analysis of cellular extracts indicated that each of four mutant lines was lacking XRCC1 protein, whereas the repair-proficient revertant line derived from one of these mutants contained a normal level of XRCC1. Although each of these cell lines expressed XRCC1 mRNA, we found in all cases a distinct point mutation resulting in crucial alterations in the encoded XRCC1 protein sequence of 633 amino acids. Two of the mutations cause non-conservative amino acid changes, Glu102→Lys and Cys390→Tyr, at positions that are invariant among hamster, mouse and human XRCC1 sequences and are located in putative functional domains. A third debilitating mutation disrupts RNA splicing, generating multiple transcripts of different length that contain deletions spanning a region of > 100 amino acids in the midsection of the XRCC1 coding sequence. A fourth mutation results in a termination codon that shortens the open reading frame to 220 amino acids, however, in the revertant cell line a further mutation in the same codon, Stop221→Leu, permits translation of a full-length functional variant protein. These mutational data indicate the importance of the putative functional regions in XRCC1, such as the BRCA1 C-terminal (BRCT) domain found in common with BRCA1 and other DNA repair and cell cycle checkpoint proteins, and also regions necessary for interaction with DNA polymerase β and DNA ligase III.

Original languageEnglish (US)
Pages (from-to)1032-1037
Number of pages6
JournalNucleic Acids Research
Volume26
Issue number4
DOIs
StatePublished - Feb 15 1998
Externally publishedYes

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DNA Repair
Cricetinae
X-Rays
Mutation
Genes
Cell Line
Amino Acids
RNA Splicing
Cell Cycle Proteins
Terminator Codon
DNA-Directed DNA Polymerase
Cricetulus
Cell Cycle Checkpoints
Point Mutation
Codon
Open Reading Frames
Amino Acid Sequence
Ovary
Messenger RNA
X-ray repair cross complementing protein 1

ASJC Scopus subject areas

  • Genetics

Cite this

Shen, M. R., Zdzienicka, M. Z., Mohrenweiser, H., Thompson, L. H., & Thelen, M. P. (1998). Mutations in hamster single-strand break repair gene XRCC1 causing defective DNA repair. Nucleic Acids Research, 26(4), 1032-1037. https://doi.org/10.1093/nar/26.4.1032

Mutations in hamster single-strand break repair gene XRCC1 causing defective DNA repair. / Shen, M. Richard; Zdzienicka, Malgorzata Z.; Mohrenweiser, Harvey; Thompson, Larry H.; Thelen, Michael P.

In: Nucleic Acids Research, Vol. 26, No. 4, 15.02.1998, p. 1032-1037.

Research output: Contribution to journalArticle

Shen, MR, Zdzienicka, MZ, Mohrenweiser, H, Thompson, LH & Thelen, MP 1998, 'Mutations in hamster single-strand break repair gene XRCC1 causing defective DNA repair', Nucleic Acids Research, vol. 26, no. 4, pp. 1032-1037. https://doi.org/10.1093/nar/26.4.1032
Shen, M. Richard ; Zdzienicka, Malgorzata Z. ; Mohrenweiser, Harvey ; Thompson, Larry H. ; Thelen, Michael P. / Mutations in hamster single-strand break repair gene XRCC1 causing defective DNA repair. In: Nucleic Acids Research. 1998 ; Vol. 26, No. 4. pp. 1032-1037.
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