| RRC ID |
35207
|
| Author |
Singh NS, Ahmad R, Sangeetha R, Varshney U.
|
| Title |
Recycling of ribosomal complexes stalled at the step of elongation in Escherichia coli.
|
| Journal |
J Mol Biol
|
| Abstract |
Translating ribosomes often stall during elongation. The stalled ribosomes are known to be recycled by tmRNA (SsrA)-mediated trans-translation. Another process that recycles the stalled ribosomes is characterized by peptidyl-tRNA release. However, the mechanism of peptidyl-tRNA release from the stalled ribosomes is not well understood. We used a defined system of an AGA-minigene containing a small open reading frame (ATG AGA AGA). Translation of the AGA-minigene mRNA is toxic to Escherichia coli because it stalls ribosomes during elongation and sequesters tRNA(Arg4) as a short-chain peptidyl-tRNA(Arg4) in the ribosomal P-site. We show that a ribosome recycling factor (RRF)-mediated process rescues the host from the AGA-minigene toxicity by releasing the peptidyl-tRNA(Arg4) from the ribosomes. The growth phenotypes of E. coli strains harboring mutant alleles of RRF and initiation factor 3 (IF3) genes and their consequences on lambdaimmP22 phage replication upon AGA-minigene expression reveal that IF3 facilitates the RRF-mediated processing of the stalled ribosomes. Additionally, we have designed a uracil DNA glycosylase gene construct, ung-stopless, whose expression is toxic to E. coli. We show that the RRF-mediated process also alleviates the ung-stopless construct-mediated toxicity to the host by releasing the ung mRNA from the ribosomes harboring long-chain peptidyl-tRNAs.
|
| Volume |
380(3)
|
| Pages |
451-64
|
| Published |
2008-7-11
|
| DOI |
10.1016/j.jmb.2008.05.033
|
| PII |
S0022-2836(08)00599-8
|
| PMID |
18565340
|
| MeSH |
Alleles
Escherichia coli / genetics*
Escherichia coli / growth & development
Escherichia coli / metabolism*
Plasmids
Prokaryotic Initiation Factor-3 / genetics
Prokaryotic Initiation Factor-3 / physiology
RNA, Bacterial / metabolism
RNA, Transfer, Amino Acyl / metabolism
Ribosomal Proteins / genetics
Ribosomal Proteins / metabolism
Ribosomes / genetics*
Ribosomes / metabolism*
|
| IF |
4.76
|
| Times Cited |
29
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Prokaryotes E. coli |
|
