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Environment dependent expression of mycobacterium hormone sensitive lipases: expression pattern under ex-vivo and individual in-vitro stress conditions in M. tuberculosis H37Ra

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Abstract

Background

Hormone-sensitive lipase (HSL) is a neutral lipase capable of hydrolysing various kinds of lipids. In comparison to single human Hormone Sensitive Lipase (hHSL), that is induced under nutritional stress, twelve serine hydrolases are annotated as HSL in Mycobacterium tuberculosis (mHSL). Mycobacterium is exposed to multiple stresses inside the host. Therefore, the present study was carried out to investigate if mHSL are also expressed under stress condition and if there is any correlation between various stress conditions and expression pattern of mHSL.

Methods and results

The expression pattern of mHSL under different environmental conditions (in-vitro and ex-vivo) were studied using qRT-PCR in M. tuberculosis H37Ra strain with 16 S rRNA as internal control. Out of 12, only two genes (lipU and lipY) were expressed at very low level in mid log phase culture under aerobic conditions, while 9 genes were expressed at stationary phase of growth. Ten mHSLs were expressed post-infection under ex-vivo conditions in time dependent manner. LipH and lipQ did not express at any time point under ex-vivo condition. The relative expression of most of the genes under individual stress was much higher than observed in ex-vivo conditions. The expression pattern of genes varied with change in stress condition.

Conclusions

Different sets of mHSL genes were expressed under different individual stress conditions pointing towards the requirement of different mHSL to combat different stress conditions. Overall, most of the mHSLs have demonstrated stress dependent expression pointing towards their role in intracellular survival of mycobacteria.

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Abbreviations

Mtb :

Mycobacterium tuberculosis

M. smegmatis :

Mycobacterium smegmatis

HSL:

Hormone Sensitive Lipases

TG:

Triglycerides

FA:

Fatty acid

RNS and ROS:

Reactive nitrogen species and Reactive oxygen species

MOI:

Multiplicity of infection

OXLDL:

Oxidized low density lipoprotein

TAG:

Triacylglycerol

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Acknowledgements

Authors would like to acknowledge Council of Scientific and Industrial Research (CSIR), New Delhi, India for financial assistance to JK {37(1609)/13/EMR-II, dated: 7/10/2013} and fellowship to SA and University Grants Commission (UGC) for providing fellowship to PS.

Funding

This study was funded by Council of Scientific and Industrial Research (CSIR), New Delhi, India (Grant no: 37(1609)/13/EMR-II).

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Authors and Affiliations

  1. Department of Biotechnology, Panjab University, BMS Block-1, Sector-25, 160014, Chandigarh, India

    Stuti Arya, Parul Singh, Jashandeep Kaur, Arbind Kumar &  Jagdeep Kaur Ph.D

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  1. Stuti Arya
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  2. Parul Singh
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  3. Jashandeep Kaur
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  5. Jagdeep Kaur Ph.D
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Contributions

Jagdeep Kaur conceived and designed the experiments. Stuti Arya performed the relevant experiments and wrote the manuscript. Parul Singh carried out hypoxia and iron stress experiments. Arbind Kumar and Jashandeep Kaur carried out bioinformatic analysis. All authors read and approved the manuscript.

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Correspondence to Jagdeep Kaur Ph.D.

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Supplementary Material 4

ESM_Fig. S1 Multiple sequence alignment of 12 mHSL: The conserved pentapeptide motif, GXSXG was shown in yellow box with red boundary; HGGG motif represented oxyanion hole was shown in green box and HGF motif was shown in blue box. These motifs are characteristics of hormone sensitive lipases

ESM_Fig. S2 Growth curve of Mtb H37Ra. Values shown are the average of three individual experiments. Error bars show standard deviations

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Arya, S., Singh, P., Kaur, J. et al. Environment dependent expression of mycobacterium hormone sensitive lipases: expression pattern under ex-vivo and individual in-vitro stress conditions in M. tuberculosis H37Ra. Mol Biol Rep 49, 4583–4593 (2022). https://doi.org/10.1007/s11033-022-07305-4

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