Molecular Cloning and Biological Characterization of the Human Excision Repair Gene ERCC-3

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 1990 Jun 1

PMID 2111438

Citations 36

Authors

G Weeda

R C Van Ham

R Masurel

A Westerveld

H Odijk

J de Wit

D BOOTSMA

A J Van Der Eb

J H Hoeijmakers

Affiliations

Soon will be listed here.

Abstract

In this report we present the cloning, partial characterization, and preliminary studies of the biological activity of a human gene, designated ERCC-3, involved in early steps of the nucleotide excision repair pathway. The gene was cloned after genomic DNA transfection of human (HeLa) chromosomal DNA together with dominant marker pSV3gptH to the UV-sensitive, incision-defective Chinese hamster ovary (CHO) mutant 27-1. This mutant belongs to complementation group 3 of repair-deficient rodent mutants. After selection of UV-resistant primary and secondary 27-1 transformants, human sequences associated with the induced UV resistance were rescued in cosmids from the DNA of a secondary transformant by using a linked dominant marker copy and human repetitive DNA as probes. From coinheritance analysis of the ERCC-3 region in independent transformants, we deduce that the gene has a size of 35 to 45 kilobases, of which one essential segment has so far been refractory to cloning. Conserved unique human sequences hybridizing to a 3.0-kilobase mRNA were used to isolate apparently full-length cDNA clones. Upon transfection to 27-1 cells, the ERCC-3 cDNA, inserted in a mammalian expression vector, induced specific and (virtually) complete correction of the UV sensitivity and unscheduled DNA synthesis of mutants of complementation group 3 with very high efficiency. Mutant 27-1 is, unlike other mutants of complementation group 3, also very sensitive toward small alkylating agents. This unique property of the mutant is not corrected by introduction of the ERCC-3 cDNA, indicating that it may be caused by an independent second mutation in another repair function. By hybridization to DNA of a human x rodent hybrid cell panel, the ERCC-3 gene was assigned to chromosome 2, in agreement with data based on cell fusion (L. H. Thompson, A. V. Carrano, K. Sato, E. P. Salazar, B. F. White, S. A. Stewart, J. L. Minkler, and M. J. Siciliano, Somat. Cell. Mol. Genet. 13:539-551, 1987).

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References

Thompson L, Busch D, Brookman K, Mooney C, Glaser D . Genetic diversity of UV-sensitive DNA repair mutants of Chinese hamster ovary cells. Proc Natl Acad Sci U S A. 1981; 78(6):3734-7. PMC: 319646. DOI: 10.1073/pnas.78.6.3734. View

Thompson L, Shiomi T, Salazar E, Stewart S . An eighth complementation group of rodent cells hypersensitive to ultraviolet radiation. Somat Cell Mol Genet. 1988; 14(6):605-12. DOI: 10.1007/BF01535314. View

Frischauf A, Lehrach H, Poustka A, Murray N . Lambda replacement vectors carrying polylinker sequences. J Mol Biol. 1983; 170(4):827-42. DOI: 10.1016/s0022-2836(83)80190-9. View

Wood R, Burki H . Repair capability and the cellular age response for killing and mutation induction after UV. Mutat Res. 1982; 95(2-3):505-14. DOI: 10.1016/0027-5107(82)90281-0. View

Hoeijmakers J, van Duin M, Westerveld A, Yasui A, BOOTSMA D . Identification of DNA repair genes in the human genome. Cold Spring Harb Symp Quant Biol. 1986; 51 Pt 1:91-101. DOI: 10.1101/sqb.1986.051.01.012. View

Naumovski L, Friedberg E . A DNA repair gene required for the incision of damaged DNA is essential for viability in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1983; 80(15):4818-21. PMC: 384136. DOI: 10.1073/pnas.80.15.4818. View

Thompson L, Brookman K, Dillehay L, Mooney C, Carrano A . Hypersensitivity to mutation and sister-chromatid-exchange induction in CHO cell mutants defective in incising DNA containing UV lesions. Somatic Cell Genet. 1982; 8(6):759-73. DOI: 10.1007/BF01543017. View

Thompson L, Salazar E, Brookman K, Collins C, Stewart S, Busch D . Recent progress with the DNA repair mutants of Chinese hamster ovary cells. J Cell Sci Suppl. 1987; 6:97-110. DOI: 10.1242/jcs.1984.supplement_6.6. View

Vermeulen W, Osseweijer P, de Jonge A, Hoeijmakers J . Transient correction of excision repair defects in fibroblasts of 9 xeroderma pigmentosum complementation groups by microinjection of crude human cell extracts. Mutat Res. 1986; 165(3):199-206. DOI: 10.1016/0167-8817(86)90055-6. View

10.

Grossman L, Caron P, Mazur S, Oh E . Repair of DNA-containing pyrimidine dimers. FASEB J. 1988; 2(11):2696-701. DOI: 10.1096/fasebj.2.11.3294078. View

11.

Sung P, Prakash L, Matson S, Prakash S . RAD3 protein of Saccharomyces cerevisiae is a DNA helicase. Proc Natl Acad Sci U S A. 1987; 84(24):8951-5. PMC: 299669. DOI: 10.1073/pnas.84.24.8951. View

12.

Higgins D, Prakash S, Reynolds P, Polakowska R, Weber S, Prakash L . Isolation and characterization of the RAD3 gene of Saccharomyces cerevisiae and inviability of rad3 deletion mutants. Proc Natl Acad Sci U S A. 1983; 80(18):5680-4. PMC: 384322. DOI: 10.1073/pnas.80.18.5680. View

13.

Stefanini M, Keijzer W, Westerveld A, BOOTSMA D . Interspecies complementation analysis of xeroderma pigmentosum and UV-sensitive Chinese hamster cells. Exp Cell Res. 1985; 161(2):373-80. DOI: 10.1016/0014-4827(85)90094-1. View

14.

Schrier P, Bernards R, Vaessen R, Houweling A, Van Der Eb A . Expression of class I major histocompatibility antigens switched off by highly oncogenic adenovirus 12 in transformed rat cells. Nature. 1983; 305(5937):771-5. DOI: 10.1038/305771a0. View

15.

Thompson L, Mooney C, Carrano A, Siciliano M . Correction of a nucleotide-excision-repair mutation by human chromosome 19 in hamster-human hybrid cells. Somat Cell Mol Genet. 1985; 11(1):87-92. DOI: 10.1007/BF01534738. View

16.

Auffray C, Rougeon F . Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980; 107(2):303-14. DOI: 10.1111/j.1432-1033.1980.tb06030.x. View

17.

Messing J, Crea R, Seeburg P . A system for shotgun DNA sequencing. Nucleic Acids Res. 1981; 9(2):309-21. PMC: 326694. DOI: 10.1093/nar/9.2.309. View

18.

Hoeijmakers J, Odijk H, Westerveld A . Differences between rodent and human cell lines in the amount of integrated DNA after transfection. Exp Cell Res. 1987; 169(1):111-9. DOI: 10.1016/0014-4827(87)90230-8. View

19.

Thompson L, Carrano A, Sato K, Salazar E, White B, Stewart S . Identification of nucleotide-excision-repair genes on human chromosomes 2 and 13 by functional complementation in hamster-human hybrids. Somat Cell Mol Genet. 1987; 13(5):539-51. DOI: 10.1007/BF01534495. View

20.

van Duin M, Vredeveldt G, Mayne L, Odijk H, Vermeulen W, Klein B . The cloned human DNA excision repair gene ERCC-1 fails to correct xeroderma pigmentosum complementation groups A through I. Mutat Res. 1989; 217(2):83-92. DOI: 10.1016/0921-8777(89)90059-1. View