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C-terminal region of Rv1039c (PPE15) protein of Mycobacterium tuberculosis targets host mitochondria to induce macrophage apoptosis

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Abstract

Mycobacterium tuberculosis (Mtb) genome possesses a unique family called Proline-Glutamate/Proline-Proline-Glutamate (PE/PPE) gene family, exclusive to pathogenic mycobacterium. Some of these proteins are known to play role in virulence and immune response modulation, but many are still uncharacterized. This study investigated the role of C-terminal region of Rv1039c (PPE15) in inducing mitochondrial perturbations and macrophage apoptosis. Our in-silico studies revealed the disordered, coiled, and hydrophobic C-terminal region in Rv1039c has similarity with C-terminal of mitochondria-targeting pro-apoptotic host proteins. Wild type Rv1039c and C-terminal deleted Rv1039c (Rv1039c-/-Cterm) recombinant proteins were purified and their M. smegmatis knock-in strains were constructed which were used for in-vitro experiments. Confocal microscopy showed localization of Rv1039c to mitochondria of PMA-differentiated THP1 macrophages; and reduced mitochondrial membrane depolarization and production of mitochondrial superoxides were observed in response to Rv1039c-/-Cterm in comparison to full-length Rv1039c. The C-terminal region of Rv1039c was found to activate caspases 3, 7 and 9 along with upregulated expression of pro-apoptotic genes like Bax and Bim. Rv1039c-/-Cterm also reduced the Cytochrome-C release from the mitochondria and the expression of AnnexinV/PI positive and TUNEL positive cells as compared to Rv1039c. Additionally, Rv1039c was observed to upregulate the TLR4-NF-κB-TNF-α signalling whereas the same was downregulated in response to Rv1039c-/-Cterm. These findings suggested that the C-terminal region of Rv1039c is a molecular mimic of pro-apoptotic host proteins which induce mitochondria-dependent macrophage apoptosis and evoke host immune response. These observations enhance our understanding about the role of PE/PPE proteins at host-pathogen interface.

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All data generated or analyzed during this study are included in this published article (and its supplementary data files). Further enquiries related to the study can be directed to the corresponding author.

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Acknowledgements

We gratefully acknowledge Professor Vikas Jain of Indian Institute of Science Education and Research [IISER], Bhopal for providing the Rv1039c-/-Cterm clone in pMSQSCHS vector.

Funding

The financial support by Department of Science and Technology [DST], Government of India [Grant Sanction no. EMR/2016/006774] and Intramural Research grant from MH, R&D cell is gratefully acknowledged. Priyanka is recipient of Senior Research Fellowship from Council of Scientific and Industrial Research [CSIR], Government of India.

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Authors and Affiliations

  1. DSKC BioDiscovery Laboratory, Department of Zoology, Miranda House, University of Delhi, Delhi, India

    Priyanka, Sadhna Sharma & Monika Sharma

  2. Department of Microbiology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India

    Mandira Varma-Basil

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  1. Priyanka
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  2. Sadhna Sharma
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Contributions

P: Experimental design, Methodology, Investigation, Formal analysis, Data curation, Writing-original draft, Writing-review and editing. SS: Supervision, Funding acquisition, Review and editing. MVB: Supervision in M. smegmatis experiments. MS: Conceptualization, Experiment design, Methodology, Supervision, Writing-review and editing of manuscript, Funding acquisition, Investigation.

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Correspondence to Monika Sharma.

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Priyanka, Sharma, S., Varma-Basil, M. et al. C-terminal region of Rv1039c (PPE15) protein of Mycobacterium tuberculosis targets host mitochondria to induce macrophage apoptosis. Apoptosis (2024). https://doi.org/10.1007/s10495-024-01965-2

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