DNA-RNA helicase activity of RAD3 protein of Saccharomyces cerevisiae

Véronique Bailly, Patrick Sung, Louise Prakash, Satya Prakash

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair of UV-damaged DNA and is essential for cell viability. The RAD3 protein exhibits a remarkable degree of sequence homology to the human excision repair protein ERCC2. The RAD3 protein is a single-stranded DNA-dependent ATPase and a DNA helicase capable of denaturing long regions of duplex DNA. Here, we demonstrate that RAD3 also possesses a potent DNA-RNA helicase activity similar in efficiency to its DNA helicase activity. The rad3 Arg-48 mutant protein, which binds but does not hydrolyze ATP, lacks the DNA-RNA unwinding activity, indicating a dependence on ATP hydrolysis. RAD3 does not show any RNA-dependent NTPase activity and, as expected, does not unwind duplex RNA. This observation suggests that RAD3 translocates on DNA in unwinding DNA-RNA duplexes. That the rad3 Arg-48 mutation inactivates the DNA and DNA-RNA helicase activities and confers a substantial reduction in the incision of UV-damaged DNA suggests a role for these activities in incision. We discuss how RAD3 helicase activities could function in tracking of DNA in search of damage sites and effect enhanced excision repair of actively transcribed genes.

Original languageEnglish (US)
Pages (from-to)9712-9716
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number21
StatePublished - 1991
Externally publishedYes

Fingerprint

RNA Helicases
DNA Helicases
Saccharomyces cerevisiae Proteins
DNA
DNA Repair
RNA
Xeroderma Pigmentosum Group D Protein
Adenosine Triphosphate
Nucleoside-Triphosphatase
Mutant Proteins
Sequence Homology
Genes
Saccharomyces cerevisiae
Cell Survival
Proteins
Hydrolysis
Mutation

Keywords

  • DNA repair/
  • Transcription/
  • Xeroderma pigmentosum

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

DNA-RNA helicase activity of RAD3 protein of Saccharomyces cerevisiae. / Bailly, Véronique; Sung, Patrick; Prakash, Louise; Prakash, Satya.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 88, No. 21, 1991, p. 9712-9716.

Research output: Contribution to journalArticle

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AU - Bailly, Véronique

AU - Sung, Patrick

AU - Prakash, Louise

AU - Prakash, Satya

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Y1 - 1991

N2 - The RAD3 gene of Saccharomyces cerevisiae is required for excision repair of UV-damaged DNA and is essential for cell viability. The RAD3 protein exhibits a remarkable degree of sequence homology to the human excision repair protein ERCC2. The RAD3 protein is a single-stranded DNA-dependent ATPase and a DNA helicase capable of denaturing long regions of duplex DNA. Here, we demonstrate that RAD3 also possesses a potent DNA-RNA helicase activity similar in efficiency to its DNA helicase activity. The rad3 Arg-48 mutant protein, which binds but does not hydrolyze ATP, lacks the DNA-RNA unwinding activity, indicating a dependence on ATP hydrolysis. RAD3 does not show any RNA-dependent NTPase activity and, as expected, does not unwind duplex RNA. This observation suggests that RAD3 translocates on DNA in unwinding DNA-RNA duplexes. That the rad3 Arg-48 mutation inactivates the DNA and DNA-RNA helicase activities and confers a substantial reduction in the incision of UV-damaged DNA suggests a role for these activities in incision. We discuss how RAD3 helicase activities could function in tracking of DNA in search of damage sites and effect enhanced excision repair of actively transcribed genes.

AB - The RAD3 gene of Saccharomyces cerevisiae is required for excision repair of UV-damaged DNA and is essential for cell viability. The RAD3 protein exhibits a remarkable degree of sequence homology to the human excision repair protein ERCC2. The RAD3 protein is a single-stranded DNA-dependent ATPase and a DNA helicase capable of denaturing long regions of duplex DNA. Here, we demonstrate that RAD3 also possesses a potent DNA-RNA helicase activity similar in efficiency to its DNA helicase activity. The rad3 Arg-48 mutant protein, which binds but does not hydrolyze ATP, lacks the DNA-RNA unwinding activity, indicating a dependence on ATP hydrolysis. RAD3 does not show any RNA-dependent NTPase activity and, as expected, does not unwind duplex RNA. This observation suggests that RAD3 translocates on DNA in unwinding DNA-RNA duplexes. That the rad3 Arg-48 mutation inactivates the DNA and DNA-RNA helicase activities and confers a substantial reduction in the incision of UV-damaged DNA suggests a role for these activities in incision. We discuss how RAD3 helicase activities could function in tracking of DNA in search of damage sites and effect enhanced excision repair of actively transcribed genes.

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