2016
DOI: 10.1074/jbc.m115.688648
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Yeast Helicase Pif1 Unwinds RNA:DNA Hybrids with Higher Processivity than DNA:DNA Duplexes

Abstract: Saccharomyces cerevisiae Pif1, an SF1B helicase, has been implicated in both mitochondrial and nuclear functions. Here we have characterized the preference of Pif1 for RNA:DNA heteroduplexes in vitro by investigating several kinetic parameters associated with unwinding. We show that the preferential unwinding of RNA:DNA hybrids is due to neither specific binding nor differences in the rate of strand separation. Instead, Pif1 is capable of unwinding RNA:DNA heteroduplexes with moderately greater processivity co… Show more

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Cited by 43 publications
(44 citation statements)
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“…The homolog of PcrA in E. coli, UvrD, has been shown to unwind RNA:DNA hybrids in vitro (Matson, 1989). In yeast, the PcrA homolog Pif1 has been shown to preferentially unwind RNA:DNA hybrids over DNA:DNA hybrids (Boulé and Zakian, 2007;Chib et al, 2016). Interestingly however, despite the established role of helicases in R-loop processing, here we find that helicases are not sufficient to process excess R-loop formation, specifically in head-on genes.…”
Section: Discussioncontrasting
confidence: 62%
“…The homolog of PcrA in E. coli, UvrD, has been shown to unwind RNA:DNA hybrids in vitro (Matson, 1989). In yeast, the PcrA homolog Pif1 has been shown to preferentially unwind RNA:DNA hybrids over DNA:DNA hybrids (Boulé and Zakian, 2007;Chib et al, 2016). Interestingly however, despite the established role of helicases in R-loop processing, here we find that helicases are not sufficient to process excess R-loop formation, specifically in head-on genes.…”
Section: Discussioncontrasting
confidence: 62%
“…However, R-loops must be problematic to some degree even in low-bias organisms such as E. coli given that modulating RNase HI levels can impact survival defects associated with rDNA inversions (Boubakri et al, 2010; Zhang et al, 2014). Interestingly, accessory helicases have been implicated in R-loop processing and conflict resolution across species (Boubakri et al, 2010; Merrikh et al, 2015; Matson, 1989; Boulé and Zakian, 2007; Chib et al, 2016), but in the case of head-on genes, our data suggest that they are not sufficient to clear the R-loops. Perhaps this is the reason for the presence of RNase HIII in organisms with higher co-orientation biases.…”
Section: Discussionmentioning
confidence: 57%
“…[31]). ScPif1 is more active on forked DNA molecules and even more active on RNA:DNA hybrids, which it unwinds efficiently even under single cycle conditions [7173]. Likewise, ScPif1 efficiently unwinds a variety of G4 structures (but not all [74]), and the rate of this unwinding is not affected measurably by the presence of a DNA trap [36,72,75].…”
Section: Biochemical Functions Of Pif1 Family Helicases Provide Himentioning
confidence: 99%