ACCURATE INITIATION OF MESSENGER-RNA SYNTHESIS IN EXTRACTS FROM SCHIZOSACCHAROMYCES-POMBE, KLUYVEROMYCES-LACTIS AND CANDIDA-GLABRATA

We demonstrate the successful adaptation to other yeast species of a p rotocol previously described for production of transcriptionally activ e whole cell extracts from Saccharomyces cerevisiae (Woontner and Jaeh ning, 1990, J. Biol. Chem. 265, 8979-8982). Extracts prepared from Sch izosaccharomyces pombe, Kluyveromyces lactis and Candida glabrata were all capable of initiating transcription from a template containing th e S. cerevisiae CYCl TATA box fused to a G-less cassette. Transcriptio n in all of the extracts was sensitive to inhibition by cl-amanitin, i ndicating that it was catalysed by RNA polymerase II, and was dramatic ally stimulated by the chimeric activator GAIA/VP16 The different extr acts used different subsets of a group of three initiation sites.