Species variation in ATP-dependent protein degradation: protease profiles differ between mycobacteria and protease functions differ between Mycobacterium smegmatis and Escherichia coli

Knipfer, Nancianne; Seth, Anjali; Roudiak, Stanislav G.; Shrader, Thomas E.

doi:10.1016/s0378-1119(99)00087-6

Cited by 22 publications

(14 citation statements)

References 26 publications

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“…The presence of a Clp-like protease system was originally proposed for M. smegmatis based on biochemical studies (29), and more recently, the completion of the M. smegmatis genome sequence ( Considering the membrane localization of DesA3, FtsH might reasonably be involved in the control of DesA3 stability. However, the degradation of DesA3 might also be controlled by either of the cytoplasmic Clp systems, as suggested by the facile degradation of soluble GFP fusions shown here.…”

Section: Resultsmentioning

confidence: 99%

In Vivo Inactivation of the Mycobacterial Integral Membrane Stearoyl Coenzyme A Desaturase DesA3 by a C-Terminus-Specific Degradation Process

et al. 2008

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DesA3 (Rv3229c) from Mycobacterium tuberculosis is a membrane-bound stearoyl coenzyme A ⌬ 9 desaturase that reacts with the oxidoreductase Rv3230c to produce oleic acid. This work provides evidence for a mechanism used by mycobacteria to regulate this essential enzyme activity. DesA3 expressed as a fusion with either a C-terminal His 6 or c-myc tag had consistently higher activity and stability than native DesA3 having the native C-terminal sequence of LAA, which apparently serves as a binding determinant for a mycobacterial protease/degradation system directed at DesA3. Fusion of only the last 12 residues of native DesA3 to the C terminus of green fluorescent protein (GFP) was sufficient to make GFP unstable. Furthermore, the comparable C-terminal sequence from the Mycobacterium smegmatis DesA3 homolog Msmeg_1886 also conferred instability to the GFP fusion. Systematic examination revealed that residues with charged side chains, large nonpolar side chains, or no side chain at the last two positions were most important for stabilizing the construct, while lesser effects were observed at the third-from-last position. Using these rules, a combinational substitution of the last three residues of DesA3 showed that either DKD or LEA gave the best enhancement of stability for the modified GFP in M. smegmatis. Moreover, upon mutagenesis of LAA at the C terminus in native DesA3 to either of these tripeptides, the modified enzyme had enhanced catalytic activity and stability. Since many proteases are conserved within bacterial families, it is reasonable that M. tuberculosis will use a similar C-terminal degradation system to posttranslationally regulate the activity of DesA3 and other proteins. Application of these rules to the M. tuberculosis genome revealed that ϳ10% the proteins encoded by essential genes may be susceptible to C-terminal proteolysis. Among these, an annotation is known for less than half, underscoring a general lack of understanding of proteins that have only temporal existence in a cell.

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Section: Resultsmentioning

confidence: 99%

In Vivo Inactivation of the Mycobacterial Integral Membrane Stearoyl Coenzyme A Desaturase DesA3 by a C-Terminus-Specific Degradation Process

et al. 2008

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“…Initially discovered in Mycobacterium tuberculosis, the bacterial ubiquitinlike tagging pathway conjugates Pup 2 to lysine side chains of protein substrates, thereby targeting them for proteasomal degradation (2). M. tuberculosis, like other actinobacterial species, contains a proteasome in addition to other smaller proteases, such as ClpXP and FtsH (3)(4)(5). Because Pup is recognized by Mpa, the proteasomal regulatory subunit, tagged (i.e.…”

mentioning

confidence: 99%

Allosteric Transitions Direct Protein Tagging by PafA, the Prokaryotic Ubiquitin-like Protein (Pup) Ligase

Ofer

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Korman

et al. 2013

Journal of Biological Chemistry

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“…Most bacteria (e.g., E. coli, B. subtilis, and Yersinia enterocolitica) possess a single clpP gene; however, some have recently been shown to contain more than one copy. For instance, Mycobacterium tuberculosis has two ClpP proteins (28) and cyanobacteria Synechocystis sp. strain PCC6803 (27) and Synechococcus sp.…”

Section: Discussionmentioning

confidence: 99%

DistinctclpPGenes Control Specific Adaptive Responses inBacillus thuringiensis

et al. 2002

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ClpP and ClpC are subunits of the Clp ATP-dependent protease, which is ubiquitous among prokaryotic and eukaryotic organisms. The role of these proteins in stress tolerance, stationary-phase adaptive responses, and virulence in many bacterial species has been demonstrated. Based on the amino acid sequences of the Bacillus subtilis clpC and clpP genes, we identified one clpC gene and two clpP genes (designated clpP1 and clpP2) in Bacillus thuringiensis. Predicted proteins ClpP1 and ClpP2 have approximately 88 and 67% amino acid sequence identity with ClpP of B. subtilis, respectively. Inactivation of clpC in B. thuringiensis impaired sporulation efficiency. The clpP1 and clpP2 mutants were both slightly susceptible to salt stress, whereas disruption of clpP2 negatively affected sporulation and abolished motility. Virulence of the clp mutants was assessed by injecting bacteria into the hemocoel of Bombyx mori larvae. The clpP1 mutant displayed attenuated virulence, which appeared to be related to its inability to grow at low temperature (25°C), suggesting an essential role for ClpP1 in tolerance of low temperature. Microscopic examination of clpP1 mutant cells grown at 25°C showed altered bacterial division, with cells remaining attached after septum formation. Analysis of lacZ transcriptional fusions showed that clpP1 was expressed at 25 and 37°C during the entire growth cycle. In contrast, clpP2 was expressed at 37°C but not at 25°C, suggesting that ClpP2 cannot compensate for the absence of ClpP1 in the clpP1 mutant cells at low temperature. Our study demonstrates that ClpP1 and ClpP2 control distinct cellular regulatory pathways in B. thuringiensis.

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Species variation in ATP-dependent protein degradation: protease profiles differ between mycobacteria and protease functions differ between Mycobacterium smegmatis and Escherichia coli

Cited by 22 publications

References 26 publications

In Vivo Inactivation of the Mycobacterial Integral Membrane Stearoyl Coenzyme A Desaturase DesA3 by a C-Terminus-Specific Degradation Process

In Vivo Inactivation of the Mycobacterial Integral Membrane Stearoyl Coenzyme A Desaturase DesA3 by a C-Terminus-Specific Degradation Process

Allosteric Transitions Direct Protein Tagging by PafA, the Prokaryotic Ubiquitin-like Protein (Pup) Ligase

DistinctclpPGenes Control Specific Adaptive Responses inBacillus thuringiensis

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