The Saccharomyces cerevisiae Sgs1 helicase efficiently unwinds G-G paired DNAs.
The Saccharomyces cerevisiae Sgs1p helicase localizes to the nucleolus and is required to maintain the integrity of the rDNA repeats. Sgs1p is a member of the RecQ DNA helicase family, which also includes Schizo-saccharomyces pombe Rqh1, and the human BLM and WRN genes. These genes encode proteins which are essential to maintenance of genomic integrity and which share a highly conserved helicase domain. Here we show that recombinant Sgs1p helicase efficiently unwinds guanine-guanine (G-G) paired DNA. Unwinding of G-G paired DNA is ATP- and Mg2+-dependent and requires a short 3' single-stranded tail. Strikingly, Sgs1p unwinds G-G paired substrates more efficiently than duplex DNAs, as measured either in direct assays or by competition experiments. Sgs1p efficiently unwinds G-G paired telomeric sequences, suggesting that one function of Sgs1p may be to prevent telomere-telomere interactions which can lead to chromosome non-disjunction. The rDNA is G-rich and has considerable potential for G-G pairing. Diminished ability to unwind G-G paired regions may also explain the deleterious effect of mutation of Sgs1 on rDNA stability, and the accelerated aging characteristic of yeast strains that lack Sgs1 as well as humans deficient in the related WRN helicase.
Sun
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Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520-8114, USA.
Bennett
R J
RJ
Maizels
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D020029:Base Pairing; D001483:Base Sequence; D004265:DNA Helicases / Q000378:metabolism*; D019175:DNA Methylation; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D053484:RecQ Helicases; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins; D014898:Werner Syndrome / Q000201:enzymology / Q000503:physiopathology
D004275:DNA, Ribosomal; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C090472:SGS1 protein, S cerevisiae; D004265:DNA Helicases; D053484:RecQ Helicases; D004264:DNA Topoisomerases, Type I