The adenylyl cyclase Rv2212 modifies the proteome and infectivity of Mycobacterium bovis BCG

Pedroza-Roldán, César; Aceves-Sánchez, Michel de Jesús; Zaveri, Anisha; Charles-Niño, Claudia; Elizondo-Quiroga, Darwin; Hernández‐Gutiérrez, Rodolfo; Allen, Kim; Visweswariah, Sandhya S.; Flores-Valdéz, Mario Alberto

doi:10.1007/s12223-014-0335-1

Cited by 6 publications

(7 citation statements)

References 27 publications

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“…Similar results have also been reported earlier for Eis protein, a thermostable aminoglycoside acetyltransferase, which enhances the survival of Mycobacterium within macrophages (13,31). A recent report by Pedroza-Roldan et al (17) has also shown that the overexpression of Rv2212 caused the enhanced replication and survival of Mycobacterium bovis BCG Pasteur 1173P2.…”

Section: Discussionsupporting

confidence: 87%

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Interaction of Erp Protein of Mycobacterium tuberculosis with Rv2212 Enhances Intracellular Survival of Mycobacterium smegmatis

et al. 2016

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The Mycobacterium tuberculosis exported repetitive protein (RvErp) is a crucial virulence-associated factor as determined by its role in the survival and multiplication of mycobacteria in cultured macrophages and in vivo. Although attempts have been made to understand the function of Erp protein, its exact role in Mycobacterium pathogenesis is still elusive. One way to determine this is by searching for novel interactions of RvErp. Using a yeast two-hybrid assay, an adenylyl cyclase (AC), Rv2212, was found to interact with RvErp. The interaction between RvErp and Rv2212 is direct and occurs at the endogenous level. The Erp protein of Mycobacterium smegmatis (MSMEG_6405, or MsErp) interacts neither with Rv2212 nor with Ms_4279, the M. smegmatis homologue of Rv2212. Deletion mutants of Rv2212 revealed its adenylyl cyclase domain to be responsible for the interaction. RvErp enhances Rv2212-mediated cyclic AMP (cAMP) production. Also, the biological significance of the interaction between RvErp and Rv2212 was demonstrated by the enhanced survival of M. smegmatis within THP-1 macrophages. Taken together, these studies address a novel mechanism by which Erp executes its function. IMPORTANCE RvErp is one of the important virulence factors of M. tuberculosis. This study describes a novel function of RvErp protein of M. tuberculosis by identifying Rv2212 as its interacting protein.Rv2212 is an adenylyl cyclase (AC) and produces cAMP, one of the prime second messengers that regulate the intracellular survival of mycobacteria. Therefore, the significance of investigating novel interactions of RvErp is paramount in unraveling the mechanisms governing the intracellular survival of mycobacteria. Discerning the molecular mechanisms used by specific mycobacterial proteins involved in infection and virulence requires an understanding of the protein-protein interaction network. The interactions of secretory proteins of Mycobacterium with the host machinery are vital for successful infection. One such secretory protein involved in virulence of Mycobacterium tuberculosis is Erp (Rv3810).The erp gene of M. tuberculosis encodes an ϳ28.0-kDa secretory protein that migrates as a 36.0-kDa protein and is present in all species of mycobacteria. Its disruption results in a marked decrease in virulence, with lower levels of survival not only in in vitro and cell culture assays but also under in vivo conditions (1, 2). It was recently shown that the nature of the erp allele strongly affects the number and the size of the lung lesions in infected animals (3). No homologue of Erp has been found in other bacterial species, making Erp a mycobacterial signature (4).Erp has a composite structure made up of three domains. While the amino-terminal domain (amino acids 1 to 80) and the carboxyterminal domain (amino acids 176 to 284) are conserved, the central domain, consisting of tandem repeats of 5 amino acids based on a PGLTS motif, is subjected to a high level of interspecies variability (1). A signal sequence is present in the amino termi...

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Section: Discussionsupporting

confidence: 87%

“…Rv2212 is known to be regulated by fatty acids and operates as an ATP sensor by integrating the signals sensing fatty acids and intracellular pH with the cAMP as the output (14). Rv2212 is also shown to modify the proteome and infectivity of Mycobacterium bovis BCG (17).…”

Section: Discussionmentioning

confidence: 99%

Interaction of Erp Protein of Mycobacterium tuberculosis with Rv2212 Enhances Intracellular Survival of Mycobacterium smegmatis

et al. 2016

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“…This latter result is in a sharp contrast with observation by Pedroza-Roldán et al ( 2015 ) concerning the BCG-Rv2212 strain, which exhibited almost identical growth with control strain in mouse organs. We suspect that this difference may be connected with different locations of Mtb and M. bovis BCG within the host cells.…”

Section: Discussioncontrasting

confidence: 98%

“…As shown earlier, M. bovis BCG with overexpressed Rv2212 did not show a significant difference in growth in vitro compared to the wild type bacteria; however, only high initial cell concentrations were used in this study. Remarkably, this strain displayed a substantial growth acceleration compared to the control strain within mouse macrophages, i.e., when stressful conditions for intracellular parasite are evident (Pedroza-Roldán et al, 2015 ). Similarly, the overexpression of Rv2212 in M. smegmatis resulted in an enhanced survival of bacteria within macrophages (Ganaie et al, 2016 ).…”

Section: Discussionmentioning

confidence: 98%

Overexpression of Adenylyl Cyclase Encoded by the Mycobacterium tuberculosis Rv2212 Gene Confers Improved Fitness, Accelerated Recovery from Dormancy and Enhanced Virulence in Mice

Shleeva

Kondratieva

Demina

et al. 2017

Front. Cell. Infect. Microbiol.

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Earlier we demonstrated that the adenylyl cyclase (AC) encoded by the MSMEG_4279 gene plays a key role in the resuscitation and growth of dormant Mycobacterium smegmatis and that overexpression of this gene leads to an increase in intracellular cAMP concentration and prevents the transition of M. smegmatis from active growth to dormancy in an extended stationary phase accompanied by medium acidification. We surmised that the homologous Rv2212 gene of M. tuberculosis (Mtb), the main cAMP producer, plays similar physiological roles by supporting, under these conditions, the active state and reactivation of dormant bacteria. To test this hypothesis, we established Mtb strain overexpressing Rv2212 and compared its in vitro and in vivo growth characteristics with a control strain. In vitro, the AC-overexpressing pMindRv2212 strain demonstrated faster growth in a liquid medium, prolonged capacity to form CFUs and a significant delay or even prevention of transition toward dormancy. AC-overexpressing cells exhibited easier recovery from dormancy. In vivo, AC-overexpressing bacteria demonstrated significantly higher growth rates (virulence) in the lungs and spleens of infected mice compared to the control strain, and, unlike the latter, killed mice in the TB-resistant strain before month 8 of infection. Even in the absence of selecting hygromycin B, all pMindRv2212 CFUs retained the Rv2212 insert during in vivo growth, strongly suggesting that AC overexpression is beneficial for bacteria. Taken together, our results indicate that cAMP supports the maintenance of Mtb cells vitality under unfavorable conditions in vitro and their virulence in vivo.

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“…Bovis enhances the survival of recombinant strain in murine macrophages (Pedroza-Roldan et al, 2015). Bioinformatic analysis study was found that cAMP receptor protein (CRP) regulated genes were conserved in pathogenic mycobacteria, and suggested that CRP regulated genes may play a role in the pathogenesis of mycobacteria (Akhter et al, 2008).…”

Section: Mycobacterium Tuberculosis Rv1265 Promotes Mycobacterial Intmentioning

confidence: 99%

Mycobacterium tuberculosisRv1265 promotes mycobacterial intracellular survival and alters cytokine profile of the infected macrophage

Luo¹,

Zeng²,

Huang³

et al. 2015

Journal of Biomolecular Structure and Dynamics

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Mycobacterium tuberculosis cAMP and underlying regulatory network are crucial for its survival and thrive in the presence of numerous stresses mounted by the host. Our studies mainly focus on the cAMP-induced M. tuberculosis gene Rv1265, which was shown to be up-regulated under hypoxia and during macrophage infection by addition of exogenous cAMP. To explore the role of Rv1265 in host-pathogen interactions, Rv1265 was expressed in a non-pathogenic Mycobacterium smegmatis. We found that Rv1265 was associated with cell envelope and can up-regulate some cell wall fatty acid components, especially the C26:0. The survival of the recombinant Ms_Rv1265 was enhanced within macrophages and under stress conditions such as low pH and SDS. Macrophages infected with Ms_Rv1265 increased transcription of pro-inflammatory cytokines IL-1β, IL-6, and IL-12 P40 and anti-inflammatory cytokine IL-10 possibly through activation of NF-κB and ERK1/2 pathway. Our findings indicate that Rv1265 can enhance mycobacterial survival within macrophages, and perturb the cytokine profile of macrophage.

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The adenylyl cyclase Rv2212 modifies the proteome and infectivity of Mycobacterium bovis BCG

Cited by 6 publications

References 27 publications

Interaction of Erp Protein of Mycobacterium tuberculosis with Rv2212 Enhances Intracellular Survival of Mycobacterium smegmatis

Interaction of Erp Protein of Mycobacterium tuberculosis with Rv2212 Enhances Intracellular Survival of Mycobacterium smegmatis

Overexpression of Adenylyl Cyclase Encoded by the Mycobacterium tuberculosis Rv2212 Gene Confers Improved Fitness, Accelerated Recovery from Dormancy and Enhanced Virulence in Mice

Mycobacterium tuberculosisRv1265 promotes mycobacterial intracellular survival and alters cytokine profile of the infected macrophage

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