Analysis of the fusA2 locus encoding EFG2 in Mycobacterium smegmatis

Seshadri, Anuradha; Samhita, Laasya; Gaur, Rahul; Malshetty, Vidyasagar; Varshney, Umesh

doi:10.1016/j.tube.2009.06.003

Cited by 14 publications

(16 citation statements)

References 43 publications

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“…Recently it has been reported that GroEL1 binds strongly to DNA and effectively functions as a DNA-protecting agent against DNase I or hydroxyl radicals (Basu et al, 2009). In addition, the orthologues of FusA2 and BcpB as well as the monooxygenase MAP0467c have been suggested to be involved in counteracting stress responses of MTB and MAP (Seshadri et al, 2009;Berney & Cook, 2010;Janagama et al, 2010;Jaeger, 2007).…”

Section: Discussionmentioning

confidence: 99%

Metabolic adaptation of Mycobacterium avium subsp. paratuberculosis to the gut environment

Weigoldt¹,

Meens²,

Bange

et al. 2013

Microbiology

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Knowledge on the proteome level about the adaptation of pathogenic mycobacteria to the environment in their natural hosts is limited. Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne's disease, a chronic and incurable granulomatous enteritis of ruminants, and has been suggested to be a putative aetiological agent of Crohn's disease in humans. Using a comprehensive LC-MS-MS and 2D difference gel electrophoresis (DIGE) approach, we compared the protein profiles of clinical strains of MAP prepared from the gastrointestinal tract of diseased cows with the protein profiles of the same strains after they were grown in vitro. LC-MS-MS analyses revealed that the principal enzymes for the central carbon metabolic pathways, including glycolysis, gluconeogenesis, the tricaboxylic acid cycle and the pentose phosphate pathway, were present under both conditions. Moreover, a broad spectrum of enzymes for boxidation of lipids, nine of which have been shown to be necessary for mycobacterial growth on cholesterol, were detected in vivo and in vitro. Using 2D-DIGE we found increased levels of several key enzymes that indicated adaptation of MAP to the host. Among these, FadE5, FadE25 and AdhB indicated that cholesterol is used as a carbon source in the bovine intestinal mucosa; the respiratory enzymes AtpA, NuoG and SdhA suggested increased respiration during infection. Furthermore higher levels of the pentose phosphate pathway enzymes Gnd2, Zwf and Tal as well as of KatG, SodA and GroEL indicated a vigorous stress response of MAP in vivo. In conclusion, our results provide novel insights into the metabolic adaptation of a pathogenic mycobacterium in its natural host.

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Section: Discussionmentioning

confidence: 99%

Metabolic adaptation of Mycobacterium avium subsp. paratuberculosis to the gut environment

Weigoldt¹,

Meens²,

Bange

et al. 2013

Microbiology

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“…The right flank amplicon of 501 bp was also cloned into the pGEM-T Easy vector and released with HindIII and BglII for its further subcloning into the same sites of pYUBudgB-LF to generate pYUBMsm-udgB : : hyg. pYUBMsm-udgB : : hyg was digested with NotI and SpeI, and a~3.2 kb fragment (udgB : : hyg) was mobilized into similarly digested pPR27 (Pelicic et al, 1997) to generate pPRMsmudgB : : hyg, which was introduced into M. smegmatis mc 2 155 or its ung 2 derivative (Venkatesh et al, 2003) by electroporation to generate the single (udgB 2 ) or double (ung 2 /udgB 2 ) knockout strains using a protocol described previously (Pelicic et al, 1997;Seshadri et al, 2009). …”

Section: Methodsmentioning

confidence: 99%

Synergistic effects of UdgB and Ung in mutation prevention and protection against commonly encountered DNA damaging agents in Mycobacterium smegmatis

et al. 2010

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The incorporation of dUMP during replication or the deamination of cytosine in DNA results in the occurrence of uracils in genomes. To maintain genomic integrity, uracil DNA glycosylases (UDGs) excise uracil from DNA and initiate the base-excision repair pathway. Here, we cloned, purified and biochemically characterized a family 5 UDG, UdgB, from Mycobacterium smegmatis to allow us to use it as a model organism to investigate the physiological significance of the novel enzyme. Studies with knockout strains showed that compared with the wild-type parent, the mutation rate of the udgB " strain was approximately twofold higher, whereas the mutation rate of a strain deficient in the family 1 UDG (ung " ) was found to be~8.4-fold higher. Interestingly, the mutation rate of the double-knockout (ung " /udgB " ) strain was remarkably high, at~19.6-fold. While CG to TA mutations predominated in the ung " and ung " /udgB " strains, AT to GC mutations were enhanced in the udgB " strain. The ung " /udgB " strain was notably more sensitive to acidified nitrite and hydrogen peroxide stresses compared with the single knockouts (ung " or udgB " ). These observations reveal a synergistic effect of UdgB and Ung in DNA repair, and could have implications for the generation of attenuated strains of Mycobacterium tuberculosis.

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“…Many organisms have a single species of EF-G that functions in both elongation and termination. However, others have two forms as observed in mammalian mitochondria [130,131]. Some forms of EF-G2 can function in translocation while others are restricted to the termination phase of translation.…”

Section: Polypeptide Chain Termination and Ribosome Recycling In Mmentioning

confidence: 99%

Mechanism of protein biosynthesis in mammalian mitochondria

Christian

Spremulli

2012

Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanism

179

143

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Protein synthesis in mammalian mitochondria produces 13 proteins that are essential subunits of the oxidative phosphorylation complexes. This review provides a detailed outline of each phase of mitochondrial translation including initiation, elongation, termination, and ribosome recycling. The roles of essential proteins involved in each phase are described. All of the products of mitochondrial protein synthesis in mammals are inserted into the inner membrane. Several proteins that may help bind ribosomes to the membrane during translation are described, although much remains to be learned about this process. Mutations in mitochondrial or nuclear genes encoding components of the translation system often lead to severe deficiencies in oxidative phosphorylation, and a summary of these mutations is provided.

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Analysis of the fusA2 locus encoding EFG2 in Mycobacterium smegmatis

Cited by 14 publications

References 43 publications

Metabolic adaptation of Mycobacterium avium subsp. paratuberculosis to the gut environment

Metabolic adaptation of Mycobacterium avium subsp. paratuberculosis to the gut environment

Synergistic effects of UdgB and Ung in mutation prevention and protection against commonly encountered DNA damaging agents in Mycobacterium smegmatis

Mechanism of protein biosynthesis in mammalian mitochondria

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