The N-terminal region of the Schizosaccharomyces pombe RecQ helicase, Rqh1p, physically interacts with Topoisomerase III and is required for Rqh1p function.
The Schizosaccharomyces pombe rqh1+ gene encodes a member of the RecQ DNA helicase family. Members of this protein family are essential for the maintenance of genetic integrity. Thus, mutations in the genes encoding the human RecQ homologues Blm, Wrn and RecQ4 cause Bloom syndrome, Werner syndrome and Rothmund-Thomson syndrome, respectively-diseases which result from genome instability. S. pombe cells that lack a functional rqh1+ gene show reduced viability and display defective chromosome segregation, particularly after UV irradiation or S-phase arrest. In this study we used an rqh1+ deletion series to show that the N-terminal portion of Rqh1 is essential for Rqh1 function. Moreover, the conserved Helicase and RNaseD C-terminal (HRDC) domain of Rqh1 also plays a role in allowing cells to tolerate exposure to DNA damaging agents and the S-phase inhibitor hydroxyurea (HU). We also demonstrate that Topoisomerase III (Top3) binds to a site within the first 322 N-terminal amino acids of Rqh1 and that this binding correlates with Rqh1 function. Genetic analysis of rqh1- top3delta mutants reveals that, in the presence of functional or partially functional Rqh1 protein, Top3 is required to maintain genome integrity and cell viability.
Ahmad
Fouzia
F
School of Biological Sciences, Michael Smith Building, University of Manchester, Oxford Road, Manchester, M13 9PT, UK. Fouzia.ahmad@manchester.ac.uk
Stewart
Elspeth
E
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D004249:DNA Damage / Q000235:genetics; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D004264:DNA Topoisomerases, Type I / Q000235:genetics / Q000378:metabolism*; D006918:Hydroxyurea / Q000633:toxicity; D047468:Immunoprecipitation; D008741:Methyl Methanesulfonate / Q000633:toxicity; D009154:Mutation / Q000235:genetics; D009841:Oligonucleotides; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D053484:RecQ Helicases; D012568:Schizosaccharomyces / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000528:radiation effects; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*; D013045:Species Specificity; D014466:Ultraviolet Rays
D009841:Oligonucleotides; D029702:Schizosaccharomyces pombe Proteins; D008741:Methyl Methanesulfonate; D000251:Adenosine Triphosphatases; C506725:RECQL protein, human; D004265:DNA Helicases; D053484:RecQ Helicases; D004264:DNA Topoisomerases, Type I; D006918:Hydroxyurea