Transcriptional elements of the yeast ribosomal protein gene CYH2.

W.F Schwindinger,J.R Warner

doi:10.1016/s0021-9258(18)45630-x

Abstract

The sequences responsible for specifying and regulating the transcription of the yeast ribosomal protein gene CYH2 have been studied using deletion analysis. We have identified a region between 235 and 260 nucleotides upstream of the transcription initiation which is necessary for transcription to occur. This region includes sequences which have been identified upstream of most yeast ribosomal protein genes. In the wild type gene the initiation of transcription occurs at several sites spread over about 15 nucleotides. Two TATA regions separated by about 40 nucleotides direct initiation to those sites. Deletion of those two TATA regions reveals a cryptic TATA which directs transcription initiation to specific sites downstream.

Highlights

The sequences responsible for specifying and regu- been implicated in the efficient transcription of the ribosomal lating the transcription of the yeast ribosomal protepinrotein genes RP39A [10] and RPL25 [11].are indicated. Secondary (As) DNA-binding gene CYH2 have been studied using deletionanalysis. protein recognizes sequences similar to HOMOL 1 and RPG
Previous studies have shown that, under conditions of bal- Yeast, Bacteria, Phage, a d Plasmids-The strain of S. cerevisiue anced growth, mRNA is synthesized at thesame rate for each used is a derivative of A364A,(ATCC 22244,14) termed 5409 MATa, of the ribosomal protein genes examined, despite the fact that urd-52,his4 his7, adel,andtyrl.The plasmids pBR322 and some genes arepresent in two copies whereas othersare present in only one copy [1].there are several conditions in which the rate of synthesis of all ribosomal proteins is regulated coordinately
When glucose is added to cultures of cells growing on ethanol, the rate of ribosomal protein synthesis increases [2] as does the concentration of YEpCYH-l(l5) and theidrerivatives were propagated in Escherichia coli C600

Summary

11 To whom correspondence should be addressed

Albert Einstein College of Medicine of Yeshiva University, 1300 Morris Park Ave., Bronx, NY 10461. A set of deletions from the 5' end of the upstream region was TC GAAAAACAGCCAAAAAACAA. Four DNA fractions with deletions in the range of 200-600 bp were subcloned into pBR322X as HindIII-. A second series of deletions was made from the 3' end of the upstream region by linearizing YEpCYH-1 with XhoI and treating it. Thirteen deletions of varying sizes were constructed throughout the upstream region (Al-Al2, A15). Hybridization Techniques-Yeast DNA was blotted by the method of Southern [24]. 0.1% sodium dodecyl sulfate for 30 min a t 25 "C, and for 30 min at described in the legend to Fig. 1B.The remaining bars indicate the. A longer primer which hybridizes to of transcripts in strains containing each of the deletions, as deterboth alleles was used instead (see "Results").

RESULTS

A12 A15 r r

DISCUSSION

I I 45