Heterologous expression of a plant small heat-shock protein enhances Escherichia coli viability under heat and cold stress.
A small heat-shock protein (sHSP) that shows molecular chaperone activity in vitro was recently purified from mature chestnut (Castanea sativa) cotyledons. This protein, renamed here as CsHSP17. 5, belongs to cytosolic class I, as revealed by cDNA sequencing and immunoelectron microscopy. Recombinant CsHSP17.5 was overexpressed in Escherichia coli to study its possible function under stress conditions. Upon transfer from 37 degrees C to 50 degrees C, a temperature known to cause cell autolysis, those cells that accumulated CsHSP17.5 showed improved viability compared with control cultures. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of cell lysates suggested that such a protective effect in vivo is due to the ability of recombinant sHSP to maintain soluble cytosolic proteins in their native conformation, with little substrate specificity. To test the recent hypothesis that sHSPs may be involved in protection against cold stress, we also studied the viability of recombinant cells at 4 degrees C. Unlike the major heat-induced chaperone, GroEL/ES, the chestnut sHSP significantly enhanced cell survivability at this temperature. CsHSP17.5 thus represents an example of a HSP capable of protecting cells against both thermal extremes. Consistent with these findings, high-level induction of homologous transcripts was observed in vegetative tissues of chestnut plantlets exposed to either type of thermal stress but not salt stress.
Soto
A
A
Departamento de Biotecnologia, Escuela Tecnica Superior Ingenieros de Montes, Universidad Politécnica de Madrid, E-28040 Madrid, Spain.
Allona
I
I
Collada
C
C
Guevara
M A
MA
Casado
R
R
Rodriguez-Cerezo
E
E
Aragoncillo
C
C
Gomez
L
L
D000595:Amino Acid Sequence; D001483:Base Sequence; D003080:Cold Temperature; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics*; D015870:Gene Expression; D017343:Genes, Plant; D006360:Heat-Shock Proteins / Q000235:genetics*; D006358:Hot Temperature; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D014197:Trees / Q000235:genetics
D017931:DNA Primers; D018076:DNA, Complementary; D018744:DNA, Plant; D006360:Heat-Shock Proteins; D010940:Plant Proteins; D011994:Recombinant Proteins