Expression of Xenopus laevis histone H5 gene in yeast.
As an approach to assess linker histone function, we engineered a cDNA encoding Xenopus laevis histone H5 (XLH5), into the yeast Saccharomyces cerevisiae, which lacks any known proteins homologous to linker histones. XLH5 cDNA when fused to the yeast GAL10 promoter and 5' untranslated region (UTR) was shown to be accurately transcribed at relatively high levels in cells harvested at mid to late log after exposure to at least 22 mM galactose. The resultant 0.95 kb XLH5 transcript reached steady state levels by approx. 2 h after galactose induction. In contrast, the product, detected by anti-XLH5 antibody, was not stably expressed until 4 h or more after induction, when no apparent growth takes place. The expression product was 27% smaller than native H5 and may have been proteolytically processed. Constitutive transcription and loss of XLH5 expression product occurred using a plasmid construct containing a 275 bp fragment of the pBR322 tetr gene inserted downstream of the GAL10 promoter. This fragment carries a putative yeast cell-type-specific upstream activation sequence.
Shwed
P S
PS
Institute of Biological Sciences, National Research Council of Canada, Ottawa.
Neelin
J M
JM
Seligy
V L
VL
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA; D015870:Gene Expression; D006657:Histones / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D014158:Transcription, Genetic; D014170:Transformation, Genetic; D014982:Xenopus laevis / Q000235:genetics*
D006657:Histones; D004247:DNA