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The mating-type-switch activating protein Sap1 is required for replication fork arrest at the rDNA of fission yeast

Otros títulosSap1p and replication fork blockage in S. pombe rDNA
AutorMejía-Ramírez, Eva; Sánchez-Gorostiaga, Alicia; Krimer, Dora B. ; Schvartzman, Jorge Bernardo ; Hernandez, Pablo
Fecha de publicaciónoct-2005
EditorAmerican Society for Microbiology
CitaciónMol Cell Biol. 2005, 25(19): 8755–8761
ResumenSchizosaccharomyces pombe rDNA contains three replication fork barriers (RFB1-3) located in the non-transcribed spacer. RFB2 and RFB3 require binding of the transcription terminator factor Reb1p to two identical recognition sequences that co-localize with these barriers. RFB1, which is the strongest of the three barriers, functions in a Reb1pindependent manner and cognate DNA-binding proteins for this barrier have not been identified yet. Here, we functionally defined RFB1 within a 78-bp sequence located near the 3’-end of the rDNA coding region. A protein that specifically binds to this sequence was purified by affinity chromatography and identified as Sap1p by mass spectrometry. Specific binding to RFB1 was confirmed by using Sap1p expressed in E. coli. Sap1p is essential for viability and is required for efficient mating-type switching. Mutations in RFB1 that precluded formation of the Sap1p-RFB1 complex systematically abolished replication barrier function, indicating that Sap1p is required for replication fork blockage at RFB1.
Descripción25 p.-5 fig.-1 tab.
Versión del editorhttp://dx.doi.org/10.1128/MCB.25.19.8755-8761.2005
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