The yeast DNA ligase gene CDC9 is controlled by six orientation specific upstream activating sequences that respond to cellular proliferation but which alone cannot mediate cell cycle regulation.
By fusing the CDC9 structural gene to the PGK upstream sequences and the CDC9 upstream to lacZ, we showed that the cell cycle expression of CDC9 is largely due to transcriptional regulation. To investigate the role of six ATGATT upstream repeats in CDC9 regulation, synthetic copies of the sequence were attached to a heterologous gene. The repeats stimulated transcription strongly and additively, but, unlike conventional yeast UAS elements, only when present in one orientation. Transcription driven by the repeats declines in cells held at START of the cell cycle or in stationary phase, as occurs with CDC9. However, the repeats by themselves cannot impart cell cycle regulation to a heterologous gene. CDC9 may therefore be controlled by an activating system operating through the repeats that is sensitive to cellular proliferation and a separate mechanism that governs the periodic expression in the cell cycle.
White
J H
JH
Laboratory of Cell Propagation, National Institute for Medical Research, The Ridgeway, Mill Hill, London, UK.
Johnson
A L
AL
Lowndes
N F
NF
Johnston
L H
LH
D001483:Base Sequence; D015152:Blotting, Northern; D002453:Cell Cycle / Q000528:radiation effects; D011088:DNA Ligases / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D012331:RNA, Fungal / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000502:physiology; D014158:Transcription, Genetic; D014466:Ultraviolet Rays; D001616:beta-Galactosidase / Q000032:analysis
D012331:RNA, Fungal; D001616:beta-Galactosidase; D011088:DNA Ligases