Isolation and characterization of specialized transducing λ phages carrying ribosomal protein genes of Escherichia coli

Kitakawa, Madoka; Blumenthal, Laura; Isono, Katsumi

doi:10.1007/bf00425846

Cited by 20 publications

(4 citation statements)

References 25 publications

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“…When IQ700 (ssyF29 aroA) was lysogenized with A serC, which carries the serC, aroA, and rpsA genes (16), the growth rate returned to normal, indicating that ssyF29 is complemented by serC (Table 2). To examine the possibility that ssyF29 is a mutation in rpsA, crude ribosomal proteins were prepared from IQ646 (ssyF29 in MC4100) as well as MC4100 (ssyF+) and analyzed by the two-dimensional gel electrophoresis system of O'Farrell (22).…”

Section: Resultsmentioning

confidence: 99%

Suppressors of the secY24 mutation: identification and characterization of additional ssy genes in Escherichia coli

Shiba¹,

Ito²,

Yura³

1986

J Bacteriol

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We previously reported (Shiba et al., J. Bacteriol. 160:696-701, 1984) the isolation and characterization of the mutation (ssy) that suppresses the protein export defect due to the secY24(Ts) mutation and causes cold-sensitive growth of Escherichia coli. This report describes more systematic isolation of ssy mutations. Among temperature-resistant revertants of the secY24 mutant, 65 mutants were found to be cold sensitive. These cold-sensitive mutations have been classified by genetic mapping. Twenty-two mutations fell into the ssyA class previously described. The remaining mutations were located at five new loci: ssyB at 9.5 min between tsx and Ion; ssyD around 3 miu; ssyE at 72.5 min near secY; ssyf at 20.5 min within rpsA; and ssyG at 69.0 min near argG. Two predominant classes, ssyA and ssyB, are probably affected in protein synthesis at the elongation step, whereas the ssyF mutant contained an altered form of ribosomal protein Si (the gene product of rpsA). These cold-sensitive ssy mutations which suppress secY24 may define genes whose function is somehow involved in the secY-dependent protein secretion mechanism. However, the existence of multiple suppressor loci makes it unlikely that all of these genes specify additional components of the export machinery. A delicate balance may exist between the systems for synthesizing and exporting proteins.Many proteins are localized to the cell envelope in Escherichia coli. The mechanism of export of these noncytoplasmic proteins has been a subject of extensive recent studies. Genetic studies have suggested that the products of several genes are involved in export of proteins across the membrane. Specifically, Beckwith and co-workers and Shiba et al. have shown that the mutations in genes secA (24,25), secB (18), and secY (29) cause a defective export of the periplasmic and outer membrane proteins, accumulating precursor molecules with unprocessed signal sequences within the cell. The correct assembly of a cytoplasmic membrane protein without cleavable signal sequence also requires the secA and sec Y gene functions (35). Alternative approaches based on extragenic suppression resulted in the identification of additional genes (3,4,7,8,15,23,28). It was expected that some of these genes may code for components of the protein export machinery.The gene secY (priA) is located within the spc ribosomal protein operon. It can mutate to different alleles showing distinct phenotypes: prlA suppressing mutations in the signal sequence (7, 30), secY24(Ts) and rplO215(Am) causing conditionally lethal phenotypes impaired in protein export (11,12,29), and prlA1012 suppressing secA51(Ts) (4).The sec Y gene product has been identified as an integral membrane protein located in the cytoplasmic membrane (1, 10), suggesting that it is a membrane component of the protein export machinery. As an approach to identifying additional -components of the export machinery and to elucidating the mode of action of the SecY protein, we isolated extragenic suppressors of secY24(Ts). We have already r...

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Section: Resultsmentioning

confidence: 99%

Suppressors of the secY24 mutation: identification and characterization of additional ssy genes in Escherichia coli

Shiba¹,

Ito²,

Yura³

1986

J Bacteriol

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“…Our novel finding of an additional target of C-terminal glutamylation, the ribosomal hibernation factor YfiA, offers an additional experimental handle with which to examine the biological and molecular functions of this modification. The association of both Ct-glutamylation target proteins with the ribosome is especially interesting, because some evidence suggests that RimK modifies S6 C-termini specifically on intact ribosomes [75, 77], and RimK is known to catalyze poly-L-glutamine formation in the absence of S6 [78]. The C-terminal amino-acid residues of YfiA resemble those of S6 only in the presence of two glutamate residues in the last two positions (DDAEAGDSEE for S6 and ANFVEEVEEE for YfiA), indicating that targeting may largely be a function of YfiA’s structural association with the ribosome rather than due to a specific sequence signal.…”

Section: Discussionmentioning

confidence: 99%

Large-scale analysis of post-translational modifications in E. coli under glucose-limiting conditions

et al. 2017

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BackgroundPost-translational modification (PTM) of proteins is central to many cellular processes across all domains of life, but despite decades of study and a wealth of genomic and proteomic data the biological function of many PTMs remains unknown. This is especially true for prokaryotic PTM systems, many of which have only recently been recognized and studied in depth. It is increasingly apparent that a deep sampling of abundance across a wide range of environmental stresses, growth conditions, and PTM types, rather than simply cataloging targets for a handful of modifications, is critical to understanding the complex pathways that govern PTM deposition and downstream effects.ResultsWe utilized a deeply-sampled dataset of MS/MS proteomic analysis covering 9 timepoints spanning the Escherichia coli growth cycle and an unbiased PTM search strategy to construct a temporal map of abundance for all PTMs within a 400 Da window of mass shifts. Using this map, we are able to identify novel targets and temporal patterns for N-terminal N α acetylation, C-terminal glutamylation, and asparagine deamidation. Furthermore, we identify a possible relationship between N-terminal N α acetylation and regulation of protein degradation in stationary phase, pointing to a previously unrecognized biological function for this poorly-understood PTM.ConclusionsUnbiased detection of PTM in MS/MS proteomics data facilitates the discovery of novel modification types and previously unobserved dynamic changes in modification across growth timepoints.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-017-3676-8) contains supplementary material, which is available to authorized users.

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“…It was ascertained that under conditions when ribosome assembly does not occur (in cells irradiated with ultraviolet light), S6 does not undergo modification [66]. The RNA-binding motif was found in RimK using bioinformatics methods [67].…”

Section: Modification Of Protein S6mentioning

confidence: 99%

Posttranslational Modifications of Ribosomal Proteins in Escherichia coli

2011

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А number of ribosomal proteins inEscherichia coliundergo posttranslational modifications. Six ribosomal proteins are methylated (S11, L3, L11, L7/L12, L16, and L33), three proteins are acetylated (S5, S18, and L7), and protein S12 is methylthiolated. Extra amino acid residues are added to protein S6. С-terminal amino acid residues are partially removed from protein L31. The functional significance of these modifications has remained unclear. These modifications are not vital to the cells, and it is likely that they have regulatory functions. This paper reviews all the known posttranslational modifications of ribosomal proteins inEscherichia coli. Certain enzymes responsible for the modifications and mechanisms of enzymatic reactions are also discussed.

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Isolation and characterization of specialized transducing λ phages carrying ribosomal protein genes of Escherichia coli

Cited by 20 publications

References 25 publications

Suppressors of the secY24 mutation: identification and characterization of additional ssy genes in Escherichia coli

Suppressors of the secY24 mutation: identification and characterization of additional ssy genes in Escherichia coli

Large-scale analysis of post-translational modifications in E. coli under glucose-limiting conditions

Posttranslational Modifications of Ribosomal Proteins in Escherichia coli

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