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Heterologous Expression of a Plant Small Heat-Shock Protein
Enhances Escherichia coli Viability under
Heat and Cold
Stress1
Alvaro Soto,
Isabel Allona,
Carmen Collada,
Maria-Angeles Guevara,
Rosa Casado,
Emilio Rodriguez-Cerezo,
Cipriano Aragoncillo, and
Luis Gomez*
Departamento de Biotecnologia, Escuela Tecnica Superior Ingenieros
de Montes, Universidad Politécnica de Madrid, E-28040 Madrid,
Spain (A.S., I.A., C.C., M.-A.G., R.C., C.A., L.G.); and Centro
Nacional de Biotecnologia-Consejo Superior de Investigaciones
Científicas, Campus de Cantoblanco, 28049 Madrid, Spain
(E.R.-C.)
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°C to 50°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°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.
1
This research was supported by grant no.
BIO96-0441 from the Ministerio de Educación y Cultura, Spain, and
by grant no. 07B-012-97 from Comunidad Autónoma de Madrid, Spain.
A.S. was the recipient of a predoctoral fellowship from the Ministerio
de Educación y Cultura, Spain.
*
Corresponding author; e-mail lgomez{at}etsi.montes.upm.es; fax
34-91-543-9557.
Plant Physiol. (1999) 120: 521-528
Copyright Clearance Center: 0032-0889/99/120//08
© 1999 American Society of Plant Physiologists
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