DNA repair gene RAD3 of S. cerevisiae is essential for transcription by RNA polymerase II

Sami N. Guzder, Hongfang Qiu, Christopher H. Sommers, Patrick Sung, Louise Prakash, Satya Prakash

Research output: Contribution to journalArticle

118 Citations (Scopus)

Abstract

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair of ultraviolet-damaged DNA and is essential for cell viability. The RAD3- encoded protein shares a high degree of homology with the human ERCC2(XPD) gene product. Mutations in XPD, besides causing the cancer-prone syndrome xeroderma pigmentosum, can also result in Cockayne's syndrome and trichothiodystrophy. To investigate the role of RAD3 in viability, we examine here the effect of a recessive, temperature-sensitive (ts) conditional lethal mutation of the gene on transcription by RNA polymerase II. Upon transfer to the restrictive temperature, the rad3-ts mutant rapidly ceases growth and poly(A)+ RNA synthesis is inhibited drastically. Messenger RNA levels of all the genes examined, HIS3, TRP3, STE2, MET19, RAD23, CDC7, CDC9 and ACT1, decline rapidly upon loss of RAD3 activity. The synthesis of heat-shock- inducible HSP26 mRNA and galactose-inducible GAL7 and GAL10 mRNAs is also drastically inhibited in the rad3-ts mutant at the restrictive temperature. The RNA polymerase II transcriptional activity in extract from the rad3- ts14 strain is thermolabile, and this in vitro transcriptional defect can be fully corrected by the addition of homogeneous RAD3 protein. These findings indicate that RAD3 protein has a direct and essential role in RNA polymerase II transcription.

Original languageEnglish (US)
Pages (from-to)91-94
Number of pages4
JournalNature
Volume367
Issue number6458
DOIs
StatePublished - Jan 6 1994

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RNA Polymerase II
DNA Repair
Saccharomyces cerevisiae
Temperature
Messenger RNA
Genes
Trichothiodystrophy Syndromes
Cockayne Syndrome
Lethal Genes
Xeroderma Pigmentosum
Mutation
Proteins
Galactose
Shock
Cell Survival
Hot Temperature
DNA
Growth
Neoplasms

ASJC Scopus subject areas

  • General

Cite this

DNA repair gene RAD3 of S. cerevisiae is essential for transcription by RNA polymerase II. / Guzder, Sami N.; Qiu, Hongfang; Sommers, Christopher H.; Sung, Patrick; Prakash, Louise; Prakash, Satya.

In: Nature, Vol. 367, No. 6458, 06.01.1994, p. 91-94.

Research output: Contribution to journalArticle

Guzder, Sami N. ; Qiu, Hongfang ; Sommers, Christopher H. ; Sung, Patrick ; Prakash, Louise ; Prakash, Satya. / DNA repair gene RAD3 of S. cerevisiae is essential for transcription by RNA polymerase II. In: Nature. 1994 ; Vol. 367, No. 6458. pp. 91-94.
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abstract = "The RAD3 gene of Saccharomyces cerevisiae is required for excision repair of ultraviolet-damaged DNA and is essential for cell viability. The RAD3- encoded protein shares a high degree of homology with the human ERCC2(XPD) gene product. Mutations in XPD, besides causing the cancer-prone syndrome xeroderma pigmentosum, can also result in Cockayne's syndrome and trichothiodystrophy. To investigate the role of RAD3 in viability, we examine here the effect of a recessive, temperature-sensitive (ts) conditional lethal mutation of the gene on transcription by RNA polymerase II. Upon transfer to the restrictive temperature, the rad3-ts mutant rapidly ceases growth and poly(A)+ RNA synthesis is inhibited drastically. Messenger RNA levels of all the genes examined, HIS3, TRP3, STE2, MET19, RAD23, CDC7, CDC9 and ACT1, decline rapidly upon loss of RAD3 activity. The synthesis of heat-shock- inducible HSP26 mRNA and galactose-inducible GAL7 and GAL10 mRNAs is also drastically inhibited in the rad3-ts mutant at the restrictive temperature. The RNA polymerase II transcriptional activity in extract from the rad3- ts14 strain is thermolabile, and this in vitro transcriptional defect can be fully corrected by the addition of homogeneous RAD3 protein. These findings indicate that RAD3 protein has a direct and essential role in RNA polymerase II transcription.",
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