In vivo genomic footprint of a yeast centromere.
We have used in vivo genomic footprinting to investigate the protein-DNA interactions within the conserved DNA elements (CDEI, CDEII, and CDEIII) in the centromere from chromosome III of the yeast Saccharomyces cerevisiae. The in vivo footprint pattern obtained from wild-type cells shows that some guanines within the centromere DNA are protected from methylation by dimethyl sulfate. These results are consistent with studies demonstrating that yeast cells contain sequence-specific centromere DNA-binding proteins. Our in vivo experiments on chromosomes with mutant centromeres show that some mutations which affect chromosome segregation also alter the footprint pattern caused by proteins bound to the centromere DNA. The results of this study provide the first fine-structure map of proteins bound to centromere DNA in living yeast cells and suggest a direct correlation between these protein-DNA interactions and centromere function.
Densmore
L
L
Department of Biochemistry, University of Massachusetts, Amherst 01003.
Payne
W E
WE
Fitzgerald-Hayes
M
M
D001483:Base Sequence; D001665:Binding Sites; D002503:Centromere; D004252:DNA Mutational Analysis; D004271:DNA, Fungal / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D008745:Methylation; D008969:Molecular Sequence Data; D011485:Protein Binding; D015347:RNA Probes; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000648:ultrastructure
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D015347:RNA Probes