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Comparative Ser/Thr/Tyr phosphoproteomics between two mycobacterial species: the fast growing Mycobacterium smegmatis and the slow growing Mycobacterium bovis BCG.

Nakedi KC, Nel AJ, Garnett S, Blackburn JM, Soares NC - Front Microbiol (2015)

Bottom Line: Moreover, our study identified a number of conserved Ser/Thr phosphorylated sites and conserved Tyr phosphorylated sites across different mycobacterial species.In particular, M. bovis BCG exponential cells exhibited a much more complex and sophisticated protein phosphorylation network regulating important cellular cycle events such as cell wall biosynthesis, elongation, cell division including immediately response to stress.The differences in the two phosphoproteomes are discussed in light of different mycobacterial growth rates.

View Article: PubMed Central - PubMed

Affiliation: Blackburn Lab, Applied Proteomics and Chemical Biology Group, Division of Medical Biochemistry, Institute of Infectious Disease and Molecular Medicine, University of Cape Town Cape Town, South Africa.

ABSTRACT
Ser/Thr/Tyr protein phosphorylation plays a critical role in regulating mycobacterial growth and development. Understanding the mechanistic link between protein phosphorylation signaling network and mycobacterial growth rate requires a global view of the phosphorylation events taking place at a given time under defined conditions. In the present study we employed a phosphopeptide enrichment and high throughput mass spectrometry-based strategy to investigate and qualitatively compare the phosphoproteome of two mycobacterial model organisms: the fast growing Mycobacterium smegmatis and the slow growing Mycobacterium bovis BCG. Cells were harvested during exponential phase and our analysis detected a total of 185 phospho-sites in M. smegmatis, of which 106 were confidently localized [localization probability (LP) = 0.75; PEP = 0.01]. By contrast, in M. bovis BCG the phosphoproteome comprised 442 phospho-sites, of which 289 were confidently localized. The percentage distribution of Ser/Thr/Tyr phosphorylation was 39.47, 57.02, and 3.51% for M. smegmatis and 35, 61.6, and 3.1% for M. bovis BCG. Moreover, our study identified a number of conserved Ser/Thr phosphorylated sites and conserved Tyr phosphorylated sites across different mycobacterial species. Overall a qualitative comparison of the fast and slow growing mycobacteria suggests that the phosphoproteome of M. smegmatis is a simpler version of that of M. bovis BCG. In particular, M. bovis BCG exponential cells exhibited a much more complex and sophisticated protein phosphorylation network regulating important cellular cycle events such as cell wall biosynthesis, elongation, cell division including immediately response to stress. The differences in the two phosphoproteomes are discussed in light of different mycobacterial growth rates.

No MeSH data available.


Related in: MedlinePlus

A histogram showing the GO molecular functions of identified phosphoproteins and respective number of phosphopeptides as predicted from their genome annotations.
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Figure 2: A histogram showing the GO molecular functions of identified phosphoproteins and respective number of phosphopeptides as predicted from their genome annotations.

Mentions: In our study we have identified nine Tyr p-sites (see Table 1), of which four were also found to be phosphorylated in M. tuberculosis (Kusebauch et al., 2014): FHA-domain-containing protein (Tyr215 and Tyr232), 60 kDa chaperonin 1 (Tyr358) and conserved membrane protein mmpS3 (Tyr70) (Figure 1B). FHA-domain-containing protein is a substrate of numerous STPKs, including PknB. Phosphorylation of FHA by PnkB has implication in cell wall synthesis with a possible involvement in mycobacterial virulence (Gupta et al., 2009). Likewise both 60 kDa Chaperonin and conserved membrane protein mmpS3 have been implicated in mycobacterial virulence (Wells et al., 2013). Based on our data, we searched for possible conservation of these peptides across other bacterial species. A sequence motif derived from 60 kDa Chaperonin Tyr358 (RQEIENSDSDYDREKLQERLA) using Seq2Logo revealed an overrepresentation of Tyr358 (Figure 2). A conserved Tyr360 residue on apparently conserved peptide (SDSDYDREKL) was found in three Gram negative pathogenic species, specifically Shigella spp., Klebsiella spp., and Salmonella spp., suggesting that this conserved Tyr phosphorylation site warrants further investigation for possible roles in bacterial pathogenesis.


Comparative Ser/Thr/Tyr phosphoproteomics between two mycobacterial species: the fast growing Mycobacterium smegmatis and the slow growing Mycobacterium bovis BCG.

Nakedi KC, Nel AJ, Garnett S, Blackburn JM, Soares NC - Front Microbiol (2015)

A histogram showing the GO molecular functions of identified phosphoproteins and respective number of phosphopeptides as predicted from their genome annotations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4389566&req=5

Figure 2: A histogram showing the GO molecular functions of identified phosphoproteins and respective number of phosphopeptides as predicted from their genome annotations.
Mentions: In our study we have identified nine Tyr p-sites (see Table 1), of which four were also found to be phosphorylated in M. tuberculosis (Kusebauch et al., 2014): FHA-domain-containing protein (Tyr215 and Tyr232), 60 kDa chaperonin 1 (Tyr358) and conserved membrane protein mmpS3 (Tyr70) (Figure 1B). FHA-domain-containing protein is a substrate of numerous STPKs, including PknB. Phosphorylation of FHA by PnkB has implication in cell wall synthesis with a possible involvement in mycobacterial virulence (Gupta et al., 2009). Likewise both 60 kDa Chaperonin and conserved membrane protein mmpS3 have been implicated in mycobacterial virulence (Wells et al., 2013). Based on our data, we searched for possible conservation of these peptides across other bacterial species. A sequence motif derived from 60 kDa Chaperonin Tyr358 (RQEIENSDSDYDREKLQERLA) using Seq2Logo revealed an overrepresentation of Tyr358 (Figure 2). A conserved Tyr360 residue on apparently conserved peptide (SDSDYDREKL) was found in three Gram negative pathogenic species, specifically Shigella spp., Klebsiella spp., and Salmonella spp., suggesting that this conserved Tyr phosphorylation site warrants further investigation for possible roles in bacterial pathogenesis.

Bottom Line: Moreover, our study identified a number of conserved Ser/Thr phosphorylated sites and conserved Tyr phosphorylated sites across different mycobacterial species.In particular, M. bovis BCG exponential cells exhibited a much more complex and sophisticated protein phosphorylation network regulating important cellular cycle events such as cell wall biosynthesis, elongation, cell division including immediately response to stress.The differences in the two phosphoproteomes are discussed in light of different mycobacterial growth rates.

View Article: PubMed Central - PubMed

Affiliation: Blackburn Lab, Applied Proteomics and Chemical Biology Group, Division of Medical Biochemistry, Institute of Infectious Disease and Molecular Medicine, University of Cape Town Cape Town, South Africa.

ABSTRACT
Ser/Thr/Tyr protein phosphorylation plays a critical role in regulating mycobacterial growth and development. Understanding the mechanistic link between protein phosphorylation signaling network and mycobacterial growth rate requires a global view of the phosphorylation events taking place at a given time under defined conditions. In the present study we employed a phosphopeptide enrichment and high throughput mass spectrometry-based strategy to investigate and qualitatively compare the phosphoproteome of two mycobacterial model organisms: the fast growing Mycobacterium smegmatis and the slow growing Mycobacterium bovis BCG. Cells were harvested during exponential phase and our analysis detected a total of 185 phospho-sites in M. smegmatis, of which 106 were confidently localized [localization probability (LP) = 0.75; PEP = 0.01]. By contrast, in M. bovis BCG the phosphoproteome comprised 442 phospho-sites, of which 289 were confidently localized. The percentage distribution of Ser/Thr/Tyr phosphorylation was 39.47, 57.02, and 3.51% for M. smegmatis and 35, 61.6, and 3.1% for M. bovis BCG. Moreover, our study identified a number of conserved Ser/Thr phosphorylated sites and conserved Tyr phosphorylated sites across different mycobacterial species. Overall a qualitative comparison of the fast and slow growing mycobacteria suggests that the phosphoproteome of M. smegmatis is a simpler version of that of M. bovis BCG. In particular, M. bovis BCG exponential cells exhibited a much more complex and sophisticated protein phosphorylation network regulating important cellular cycle events such as cell wall biosynthesis, elongation, cell division including immediately response to stress. The differences in the two phosphoproteomes are discussed in light of different mycobacterial growth rates.

No MeSH data available.


Related in: MedlinePlus