Isolation and sequence of HSP30, a yeast heat-shock gene coding for a hydrophobic membrane protein

Abstract

Using differential hybridization, a gene preferentially expressed during entry into stationary phase has been isolated. Subsequent analysis indicated that this gene corresponds to a heat-shock gene. The nucleotide sequence has been determined. It revealed a 332 amino-acid protein. No similarities to any previously known protein have been noted. The protein is very hydrophobic and is predicted to have a membraneous localisation. In agreement with this hypothesis, the analysis of membrane proteins from stationary-phase cells showed that a strain carrying this gene on a multicopy vector overproduces a protein of 30 kDa. This protein was recognized by antibodies directed against the N-terminal portion of the gene product. Considering its induction in response to heat shock and the apparent molecular weight of its product, this gene was designated HSP30.

Stress induction of HSP30, the plasma membrane heat shock protein gene of Saccharomyces cerevisiae, appears not to use known stress-regulated transcription factors

“…Previous work has shown that Hsp30 mRNA is induced during entry to stationary phase under conditions where glucose becomes limiting (Regnacq & Boucherie, 1992 ;Riou et al, 1997) ; also that Hsp30 protein is induced in purified plasma membranes by glucose limitation, heat shock and exposure to either ethanol or sorbic acid (Panaretou & Piper, 1992;Regnacq & Boucherie, 1992;Piper et al, 1994Piper et al, , 1997. The effects of different stresses on Hsp30 mRNA levels are shown in Fig.…”

“…Unexpectedly our data indicate that stress activation of this gene is due to neither of these promoter elements. HSP30 is induced by more than one stress, including several that also induce the STRE [heat shock, ethanol, sorbate exposure (Figs 1 and 3-6) and carbon starvation (Regnacq & Boucherie, 1992; Riou et al, 1997)l. However, it is not induced by certain other STRE-inducing stresses (moderate osmostress, nitrogen starvation). Moreover the basal and stress-induced expressions of this gene, unlike those of the STRE and PDSE, appear to be largely independent of PKA activity (Fig.…”

“…by ligation of the 2.4 kb NdeI fragment of pUC19HSP30 (Regnacq & Boucherie, 1992) into NdeI-cleaved YIplac204 (Geitz & Sugino, 1988). YIp30 contains the entire HSP30 ORF and 963 bp of upstream DNA, together with a unique EcoRV site within the TRPI gene of the vector.…”

The HTL1 gene (YCR020W‐b) of Saccharomyces cerevisiae is necessary for growth at 37°C, and for the conservation of chromosome stability and fertility

“…The 10 complementing clones share a common sequence, containing what appeared to be a single 'significant' (i.e. >300 bp) putative ORF corresponding to HSP30 (a membrane ATPase) (Panaretou and Piper, 1992;Regnacq and Boucherie, 1993), but are missing either MAK31 or MAK32 or both (data not shown). The smallest complementing subclone (pRC2) derived from the original y12 kb complementing fragments contained an approximately 2.5 kb BamHI-NruI fragment, including the 1 kb HSP30 ORF.…”

Structure of Genes for Hsp30 from the White-rot fungusCoriolus versicolorand the Increase of their Expression by Heat Shock and Exposure to a Hazardous Chemical

“…7,8) In this report, we describe the structure of two genomic genes for Hsp30 from C. versicolor and their promoter regions.…”