Identification and purification of a factor that binds to the Mlu I cell cycle box of yeast DNA replication genes.
In Saccharomyces cerevisiae, the genes encoding at least 10 enzymes involved in DNA replication are periodically expressed in the late G1 and S phases of the cell cycle. All of these genes have one copy or more of the sequence ACGCGT, which conforms to the recognition site for the Mlu I restriction endonuclease. For the CDC21, CDC9, and POL1 genes, the Mlu I site has been shown to be absolutely required for periodic transcription. Using nuclear extracts fractionated by conventional and oligonucleotide affinity chromatography, we have purified a 17-kDa protein that recognizes the Mlu I motif. Synthetic oligonucleotides containing mutated Mlu I sites do not bind the protein. In contrast, synthetic oligonucleotides derived from the CDC2, CDC6, and CDC21 genes, which are expressed with the same timing as POL1, bind purified protein efficiently.
Verma
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R
Braun Laboratories, California Institute of Technology, Pasaden 91125.
Patapoutian
A
A
Gordon
C B
CB
Campbell
J L
JL
D001426:Bacterial Proteins; D001483:Base Sequence; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002453:Cell Cycle / Q000235:genetics; D002467:Cell Nucleus / Q000502:physiology; D004256:DNA Polymerase I / Q000235:genetics; D004261:DNA Replication; D003850:Deoxyribonuclease I; D015252:Deoxyribonucleases, Type II Site-Specific; D005656:Fungal Proteins / Q000235:genetics; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D002352:Carrier Proteins; D005656:Fungal Proteins; D015345:Oligonucleotide Probes; D004256:DNA Polymerase I; C035041:endodeoxyribonuclease MluI; D003850:Deoxyribonuclease I; D015252:Deoxyribonucleases, Type II Site-Specific