Mutational analysis of the Escherichia coli DEAD box protein CsdA

被引:36
|
作者
Turner, Anne-Marie W.
Love, Cheraton F.
Alexander, Rebecca W.
Jones, Pamela G. [1 ]
机构
[1] Winston Salem State Univ, Dept Life Sci, Winston Salem, NC 27110 USA
[2] Wake Forest Univ, Dept Chem, Winston Salem, NC 27109 USA
关键词
DEPENDENT RNA HELICASE; TRANSLATION INITIATION; POLYNUCLEOTIDE PHOSPHORYLASE; SACCHAROMYCES-CEREVISIAE; TERMINAL DOMAIN; SHOCK PROTEIN; BINDING; FAMILY; GENE; ADAPTATION;
D O I
10.1128/JB.01509-06
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The Escherichia coli cold shock protein CsdA is a member of the DEAD box family of ATP-dependent RNA helicases, which share a core of nine conserved motifs. The DEAD (Asp-Glu-Ala-Asp) motif for which this family is named has been demonstrated to be essential for ATP hydrolysis. We show here that CsdA exhibits in vitro ATPase and helicase activities in the presence of short RNA duplexes with either 3' or 5' extensions at 15 degrees C. In contrast to wild-type CsdA, a DQAD variant of CsdA (Glu-157 -> Gln) had no detectible helicase or ATPase activity at 15 degrees C in vitro. A plasmid encoding the DQAD variant was also unable to suppress the impaired growth of the csdA null mutant at 15 degrees C. Plasmid-encoded CsdA Delta 444, which lacks most of the carboxy-terminal extension, enhanced the growth of a csdA null mutant at 25 degrees C but not at 15 degrees C; this truncated protein also has limited in vitro activity at 15 degrees C. These results support the physiological function of CsdA as a DEAD box ATP-dependent RNA helicase at low temperature.
引用
收藏
页码:2769 / 2776
页数:8
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