Gene- and Strand-specific Repair in Vitro: Partial Purification of a Transcription-repair Coupling Factor

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1991 Sep 15

PMID 1896474

Citations 39

Authors

C P Selby

A Sancar

Affiliations

Soon will be listed here.

Abstract

In eukaryotic and prokaryotic cells, activity transcribed genes and, in some instances, the template strand of these genes have been found to be repaired 2-10 times more rapidly than nontranscribed genes or the coding strand of transcribed genes. We demonstrate here gene- and template strand-specific repair synthesis in vitro by using an Escherichia coli cell-free extract and a plasmid carrying a gene with the strong tac promoter. Strand-specific repair of UV, 4'-hydroxymethyl1-4,5',8-trimethylpsoralen, and cis-dicholorodiammine platinum(II) damage was dependent upon transcription and a functional nucleotide excision repair system and was stimulated by 6% (wt/vol) polyethylene glycol. A defined system consisting of the transcription and repair proteins in highly purified form did not perform strand-specific repair; however, active fractions of extract conferred strand specificity to the defined system. Transcription-repair coupling activity was partially purified from extract by successive DEAE-agarose and gel filtration chromatography. The coupling factor is heat-labile, with an estimated Mr of 100,000.

Citing Articles

In vitro DNA repair genomics using XR-seq with and mammalian cell-free extracts.

Cao X, Kose C, Selby C, Sancar A Proc Natl Acad Sci U S A. 2023; 120(43):e2314233120.

PMID: 37844222 PMC: 10614213. DOI: 10.1073/pnas.2314233120.

The Transcription-Repair Coupling Factor Mfd Prevents and Promotes Mutagenesis in a Context-Dependent Manner.

Lindsey-Boltz L, Sancar A Front Mol Biosci. 2021; 8:668290.

PMID: 34095223 PMC: 8174841. DOI: 10.3389/fmolb.2021.668290.

A Nobel-Winning Scientist: Aziz Sancar and the Impact of his Work on the Molecular Pathology of Neoplastic Diseases.

Pehlivanoglu B, Aysal A, Kececi S, Ekmekci S, Erdogdu I, Ertunc O Turk Patoloji Derg. 2021; 37(2):93-105.

PMID: 33973640 PMC: 10512686. DOI: 10.5146/tjpath.2020.01504.

Structural basis for transcription complex disruption by the Mfd translocase.

Kang J, Llewellyn E, Chen J, Olinares P, Brewer J, Chait B Elife. 2021; 10.

PMID: 33480355 PMC: 7864632. DOI: 10.7554/eLife.62117.

A Peek Inside the Machines of Bacterial Nucleotide Excision Repair.

Kraithong T, Hartley S, Jeruzalmi D, Pakotiprapha D Int J Mol Sci. 2021; 22(2).

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References

Venema J, Mullenders L, Natarajan A, van Zeeland A, Mayne L . The genetic defect in Cockayne syndrome is associated with a defect in repair of UV-induced DNA damage in transcriptionally active DNA. Proc Natl Acad Sci U S A. 1990; 87(12):4707-11. PMC: 54186. DOI: 10.1073/pnas.87.12.4707. View

Weeda G, Van Ham R, Vermeulen W, BOOTSMA D, Van Der Eb A, Hoeijmakers J . A presumed DNA helicase encoded by ERCC-3 is involved in the human repair disorders xeroderma pigmentosum and Cockayne's syndrome. Cell. 1990; 62(4):777-91. DOI: 10.1016/0092-8674(90)90122-u. View

Fuller R, Kaguni J, Kornberg A . Enzymatic replication of the origin of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1981; 78(12):7370-4. PMC: 349268. DOI: 10.1073/pnas.78.12.7370. View

Lu A, Clark S, Modrich P . Methyl-directed repair of DNA base-pair mismatches in vitro. Proc Natl Acad Sci U S A. 1983; 80(15):4639-43. PMC: 384099. DOI: 10.1073/pnas.80.15.4639. View

Sancar G, Sancar A, RUPP W . Sequences of the E. coli uvrC gene and protein. Nucleic Acids Res. 1984; 12(11):4593-608. PMC: 318860. DOI: 10.1093/nar/12.11.4593. View

Lindahl T . DNA repair enzymes. Annu Rev Biochem. 1982; 51:61-87. DOI: 10.1146/annurev.bi.51.070182.000425. View

Sancar A, RUPP W . A novel repair enzyme: UVRABC excision nuclease of Escherichia coli cuts a DNA strand on both sides of the damaged region. Cell. 1983; 33(1):249-60. DOI: 10.1016/0092-8674(83)90354-9. View

Amann E, Brosius J, Ptashne M . Vectors bearing a hybrid trp-lac promoter useful for regulated expression of cloned genes in Escherichia coli. Gene. 1983; 25(2-3):167-78. DOI: 10.1016/0378-1119(83)90222-6. View

Meneghini R, Menck C, Schumacher R . Mechanisms of tolerance to DNA lesions in mammalian cells. Q Rev Biophys. 1981; 14(3):381-432. DOI: 10.1017/s0033583500002353. View

10.

SAUERBIER W, Hercules K . Gene and transcription unit mapping by radiation effects. Annu Rev Genet. 1978; 12:329-63. DOI: 10.1146/annurev.ge.12.120178.001553. View

11.

Wickner W, Brutlag D, Schekman R, Kornberg A . RNA synthesis initiates in vitro conversion of M13 DNA to its replicative form. Proc Natl Acad Sci U S A. 1972; 69(4):965-9. PMC: 426605. DOI: 10.1073/pnas.69.4.965. View

12.

RUPP W, Wilde 3rd C, Reno D, HOWARD-FLANDERS P . Exchanges between DNA strands in ultraviolet-irradiated Escherichia coli. J Mol Biol. 1971; 61(1):25-44. DOI: 10.1016/0022-2836(71)90204-x. View

13.

Michalke H, Bremer H . RNA synthesis in Escherichia coli after irradiation with ultraviolet light. J Mol Biol. 1969; 41(1):1-23. DOI: 10.1016/0022-2836(69)90122-3. View

14.

Selby C, Sancar A . Transcription preferentially inhibits nucleotide excision repair of the template DNA strand in vitro. J Biol Chem. 1990; 265(34):21330-6. View

15.

Wood R . DNA repair. Seven genes for three diseases. Nature. 1991; 350(6315):190. DOI: 10.1038/350190a0. View

16.

McGregor W, Chen R, LUKASH L, Maher V, McCormick J . Cell cycle-dependent strand bias for UV-induced mutations in the transcribed strand of excision repair-proficient human fibroblasts but not in repair-deficient cells. Mol Cell Biol. 1991; 11(4):1927-34. PMC: 359877. DOI: 10.1128/mcb.11.4.1927-1934.1991. View

17.

Chen R, Maher V, McCormick J . Effect of excision repair by diploid human fibroblasts on the kinds and locations of mutations induced by (+/-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10- tetrahydrobenzo[a]pyrene in the coding region of the HPRT gene. Proc Natl Acad Sci U S A. 1990; 87(21):8680-4. PMC: 55021. DOI: 10.1073/pnas.87.21.8680. View

18.

Mellon I, HANAWALT P . Induction of the Escherichia coli lactose operon selectively increases repair of its transcribed DNA strand. Nature. 1989; 342(6245):95-8. DOI: 10.1038/342095a0. View

19.

Sancar A, Sancar G . DNA repair enzymes. Annu Rev Biochem. 1988; 57:29-67. DOI: 10.1146/annurev.bi.57.070188.000333. View

20.

Liu L, Wang J . Supercoiling of the DNA template during transcription. Proc Natl Acad Sci U S A. 1987; 84(20):7024-7. PMC: 299221. DOI: 10.1073/pnas.84.20.7024. View