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MBC in Press, published online ahead of print March 21, 2002
Mol. Biol. Cell 10.1091/mbc.02-02-0021

A more recent version of this article appeared on June 1, 2002
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Submitted on November 13, 2001
Revised on January 28, 2002
Accepted on February 22, 2002

Schizosaccharomyces pombe pfh1+ encodes an essential 5' to 3' DNA helicase that is a member of the PIF1 sub-family of DNA helicases

Jin-Qiu Zhou1, Haiyan Qi1, Vincent P. Schulz1, Maria K. Mateyak1, Ellen K. Monson1, and Virginia A. Zakian1*

1 Princeton University, Department of Molecular Biology, Princeton, NJ 08544

* Corresponding author. E-mail address: vzakian{at}molbio.princeton.edu.

The Saccharomyces cerevisiae Pif1p DNA helicase is the prototype member of a helicase sub-family conserved from yeast to humans. S. cerevisiae has two PIF1-like genes, PIF1 itself and RRM3, that have roles in maintenance of telomeric, ribosomal, and mitochondrial DNA. Here we describe the isolation and characterization of pfh1+, a Schizosaccharomyces pombe gene that encodes a Pif1-like protein. Pfh1p was the only S. pombe protein with high identity to Saccharomyces Pif1p. Unlike the two S. cerevisiae Pif1 sub-family proteins, the S. pombe Pfh1p was essential. Like Saccharomyces Pif1p, a truncated form of the S. pombe protein had 5' to 3' DNA helicase activity. Point mutations in an invariant lysine residue in the ATP binding pocket of Pfh1p had the same phenotype as deleting pfh1+, demonstrating that the ATPase/helicase activity of Pfh1p was essential. Although mutant spores depleted for Pfh1p proceeded through S phase, they arrested with a terminal cellular phenotype consistent with a post initiation defect in DNA replication. Telomeric DNA was modestly shortened in the absence of Pfh1p. However, genetic analysis demonstrated that maintenance of telomeric DNA was not the sole essential function of S. pombe Pfh1p.




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