Journal
JOURNAL OF MOLECULAR BIOLOGY
Volume 332, Issue 3, Pages 575-584Publisher
ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD
DOI: 10.1016/S0022-2836(03)00953-7
Keywords
RbfA; 16 S rRNA processing; ribosome; cold shock
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Funding
- NIGMS NIH HHS [GM19043] Funding Source: Medline
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RbfA, a 30 S ribosome-binding factor, is a multicopy suppressor of a cold-sensitive C23U mutation of the 16 S rRNA and is required for efficient processing of the 16 S rRNA. At 37 degreesC, DeltarbfA cells show accumulation of ribosomal subunits and 16 S rRNA precursor with a significantly reduced polysome profile in comparison with wild-type cells. RbfA is also a cold-shock protein essential for Escherichia coli cells to adapt to low temperature. In this study, we examined its association with the ribosome and its role in 16 S rRNA processing and ribosome profiles at low temperature. In wild-type cells, following cold shock at 15 degreesC, the amount of free RbfA remained largely stable, while that of its 30 S subunit-associated form became several times greater than that at 37 degreesC and a larger fraction of total 30 S subunits was detected to be RbfA-containing. In DeltarbfA cells, the pre-16 S rRNA amount increased after cold shock with a concomitant reduction of the mature 16 S rRNA amount and the formation of polysomes was further reduced. A closer examination revealed that 30 S ribosomal subunits of DeltarbfA cells at low temperature contained primarily pre-16 S rRNA and little mature 16 S rRNA. Our results indicate that the cold sensitivity of DeltarbfA cells is directly related to their lack of translation initiation-capable 30 S subunits containing mature 16 S rRNA at low temperature. Importantly, when the C-terminal 25 residue sequence was deleted, the resulting RbfADelta25 lost the abilities to stably associate with the 30 S subunit and to suppress the dominant-negative, cold-sensitive phenotype of the C23U mutation in 16 S rRNA but was able to suppress the 16 S rRNA processing defect and the cold-sensitive phenotype of the DeltarbfA cells, suggesting that RbfA may interact with the 30 S ribosome at more than one site or function in more than one fashion in assisting the 16 S rRNA maturation at low temperature. (C) 2003 Elsevier Ltd. All rights reserved.
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