Northern blot hybridization analysis of a series of 5' end, 3' end and internal deletions has revealed that at least four different regions are involved in the regulation of the expression of TIP1, a cold shock-inducible gene of Saccharomyces cerevisiae. One of these four regions has negative effect on the expression of the TIP1 gene, while the others are responsible for the activation and cold shock-induction of the gene. A fragment involved in the cold-shock induction of TIP1 was used as a probe in gel retardation assays to identify the cold shock-factor. The cold shock-factor could be detected in cells grown at 30 degrees C as well as 10 degrees C, but both the amount of the factor and its affinity to DNA were found to increase 2-3-fold after cold shock. In addition, another factor was found to bind just upstream of the cold shock element, in a region where a transcriptional activator was predicted to function by Northern blot hybridization analysis. The amount of this activating factor and its affinity for DNA was not affected by temperature. Implications of our data on possible mechanisms of transcriptional regulation of the TIP1 gene by cold shock are discussed.
Base Sequence Blotting, Northern