Effects of heterologous expression of CspB, the major cold shock protein of Bacillus subtilis, on protein synthesis in Escherichia coli

1997 ◽  
Vol 253 (6) ◽  
pp. 745-752 ◽  
Author(s):  
P. Graumann ◽  
M. A. Marahiel
Keyword(s):  
Escherichia Coli ◽  
Protein Synthesis ◽  
Bacillus Subtilis ◽  
Heterologous Expression ◽  
Cold Shock ◽  
Cold Shock Protein

scholarly journals Major cold shock protein of Escherichia coli.

1990 ◽  
Vol 87 (1) ◽  
pp. 283-287 ◽  
Author(s):  
J. Goldstein ◽  
N. S. Pollitt ◽  
M. Inouye
Keyword(s):  
Escherichia Coli ◽  
Cold Shock ◽  
Cold Shock Protein

scholarly journals Localization of Cold Shock Proteins to Cytosolic Spaces Surrounding Nucleoids in Bacillus subtilis Depends on Active Transcription

2001 ◽  
Vol 183 (21) ◽  
pp. 6435-6443 ◽  
Author(s):  
Michael H. W. Weber ◽  
Arsen V. Volkov ◽  
Ingo Fricke ◽  
Mohamed A. Marahiel ◽  
Peter L. Graumann
Keyword(s):  
Bacillus Subtilis ◽  
Cold Shock ◽  
Fluorescent Protein ◽  
Cold Shock Protein ◽  
Initiation Of Translation ◽  
Specific Localization ◽  
Active Transcription ◽  
Green Fluorescent ◽  
Shock Proteins ◽  
Mutant Cells

ABSTRACT 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 β-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 incspB cspC and cspB cspDdouble-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.

scholarly journals Crystal structure of CspA, the major cold shock protein of Escherichia coli.

1994 ◽  
Vol 91 (11) ◽  
pp. 5119-5123 ◽  
Author(s):  
H. Schindelin ◽  
W. Jiang ◽  
M. Inouye ◽  
U. Heinemann
Keyword(s):  
Crystal Structure ◽  
Escherichia Coli ◽  
Cold Shock ◽  
Cold Shock Protein

scholarly journals Mutation Analysis of the 5′ Untranslated Region of the Cold Shock cspA mRNA of Escherichia coli

1999 ◽  
Vol 181 (20) ◽  
pp. 6284-6291 ◽  
Author(s):  
Kunitoshi Yamanaka ◽  
Masanori Mitta ◽  
Masayori Inouye
Keyword(s):  
Escherichia Coli ◽  
Untranslated Region ◽  
Cold Shock ◽  
Cold Shock Protein ◽  
Translation Efficiency ◽  
Negative Effect ◽  
Downstream Box ◽  
The Stability ◽  
The Relationship ◽  
Positive Effect

ABSTRACT The mRNA for CspA, a major cold shock protein in Escherichia coli, contains an unusually long (159 bases) 5′ untranslated region (5′-UTR), and its stability has been shown to play a major role in cold shock induction of CspA. The 5′-UTR of the cspAmRNA has a negative effect on its expression at 37°C but has a positive effect upon cold shock. In this report, a series ofcspA-lacZ fusions having a 26- to 32-base deletion in the 5′-UTR were constructed to examine the roles of specific regions within the 5′-UTR in cspA expression. It was found that none of the deletion mutations had significant effects on the stability of mRNA at both 37 and 15°C. However, two mutations (Δ56-86 and Δ86-117) caused a substantial increase of β-galactosidase activity at 37°C, indicating that the deleted regions contain a negativecis element(s) for translation. A mutation (Δ2-27) deleting the highly conserved cold box sequence had little effect on cold shock induction of β-galactosidase. Interestingly, three mutations (Δ28-55, Δ86-117, and Δ118-143) caused poor cold shock induction of β-galactosidase. In particular, the Δ118-143 mutation reduced the translation efficiency of the cspA mRNA to less than 10% of that of the wild-type construct. The deleted region contains a 13-base sequence named upstream box (bases 123 to 135), which is highly conserved in cspA, cspB,cspG, and cspI, and is located 11 bases upstream of the Shine-Dalgarno (SD) sequence. The upstream box might be another cis element involved in translation efficiency of the cspA mRNA in addition to the SD sequence and the downstream box sequence. The relationship between the mRNA secondary structure and translation efficiency is discussed.

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