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Journal of Bacteriology, November 2001, p. 6435-6443, Vol. 183, No. 21
Biochemie, Fachbereich Chemie,
Philipps-Universität Marburg, 35032 Marburg, Germany
Received 18 June 2001/Accepted 21 June 2001
Using immunofluorescence microscopy and a fusion of a cold shock
protein (CSP), CspB, to green fluorescent protein (GFP), we showed that
in growing cells Bacillus subtilis CSPs specifically localize to cytosolic regions surrounding the nucleoid. The
subcellular localization of CSPs is influenced by the structure of
the nucleoid. Decondensed chromosomes in smc mutant
cells reduced the sizes of the regions in which CSPs localized, while
cold shock-induced chromosome compaction was accompanied by an
expansion of the space in which CSPs were present. As a control,
histone-like protein HBsu localized to the nucleoids, while
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.21.6435-6443.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Localization of Cold Shock Proteins to Cytosolic Spaces
Surrounding Nucleoids in Bacillus subtilis Depends on
Active Transcription
-galactosidase and GFP were detectable throughout the cell.
After inhibition of translation, CspB-GFP was still present
around the nucleoids in a manner similar to that in cold-shocked cells.
However, in stationary-phase cells and after inhibition of
transcription, CspB was distributed throughout the cell, indicating
that specific localization of CspB depends on active transcription and
is not due to simple exclusion from the nucleoid. Furthermore, we
observed that nucleoids are more condensed and frequently abnormal in
cspB cspC and cspB cspD
double-mutant cells. This suggests that the function of CSPs affects
chromosome structure, probably through coupling of transcription to
translation, which is thought to decondense nucleoids. In addition, we
found that cspB cspD and cspB cspC double
mutants are defective in sporulation, with a block at or before stage
0. Interestingly, CspB and CspC are depleted from the forespore
compartment but not from the mother cell. In toto, our findings
suggest that CSPs localize to zones of newly synthesized RNA,
coupling transcription with initiation of translation.
*
Corresponding author. Mailing address: Biochemie,
Fachbereich Chemie, Hans-Meerwein-Straße, Philipps-Universität
Marburg, 35032 Marburg, Germany. Phone: 49 (0) 6421 2825539. Fax: 49 (0) 6421 2822191. E-mail:
graumann{at}chemie.uni-marburg.de.
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