Rtf1-mediated eukaryotic site-specific replication termination.
The molecular mechanisms mediating eukaryotic replication termination and pausing remain largely unknown. Here we present the molecular characterization of Rtf1 that mediates site-specific replication termination at the polar Schizosaccharomyces pombe barrier RTS1. We show that Rtf1 possesses two chimeric myb/SANT domains: one is able to interact with the repeated motifs encoded by the RTS1 element as well as the elements enhancer region, while the other shows only a weak DNA binding activity. In addition we show that the C-terminal tail of Rtf1 mediates self-interaction, and deletion of this tail has a dominant phenotype. Finally, we identify a point mutation in Rtf1 domain I that converts the RTS1 element into a replication barrier of the opposite polarity. Together our data establish that multiple protein DNA and protein-protein interactions between Rtf1 molecules and both the repeated motifs and the enhancer region of RTS1 are required for site-specific termination at the RTS1 element.
Eydmann
T
T
Marie Curie Research Institute, The Chart, Oxted RH8 0TL, United Kingdom.
Sommariva
E
E
Inagawa
T
T
Mian
S
S
Klar
A J S
AJ
Dalgaard
J Z
JZ
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D004247:DNA / Q000737:chemistry; D004261:DNA Replication; D004275:DNA, Ribosomal / Q000737:chemistry; D004742:Enhancer Elements, Genetic; D005656:Fungal Proteins / Q000737:chemistry; D008957:Models, Genetic; D008969:Molecular Sequence Data; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D012568:Schizosaccharomyces / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D035181:TATA-Box Binding Protein / Q000235:genetics*; D020798:Two-Hybrid System Techniques
D004275:DNA, Ribosomal; D005656:Fungal Proteins; C107273:RTF1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D035181:TATA-Box Binding Protein; D004247:DNA