Jitender S Verma1, Yash Gupta2, Deepthi Nair3, Nikhat Manzoor4, Rajinder S Rautela5, Arvind Rai5, Vishwa M Katoch6. 1. Department of Orthopaedics, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi 110029, India jitu_jitender1@rediffmail.com. 2. National JALMA Institute for Leprosy and Other Mycobacterial Diseases (ICMR), Tajganj, Agra 282001, India. 3. Department of Microbiology, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi 110029, India. 4. Department of Bio-Sciences, Jamia Millia Islamia, New Delhi 110025, India. 5. National Centre for Disease Control, Sham Nath Marg, New Delhi 110054, India. 6. Director General, Indian Council of Medical Research and Secretary, Department of Health Research, Ministry of Health and Family Welfare, India.
Abstract
OBJECTIVES: To evaluate gidB alterations for possible impact on the cumulative mechanism underlying the acquisition of high-level streptomycin resistance in Mycobacterium tuberculosis. METHODS: Fifty-two isolates with high streptomycin resistance and 23 isolates with low streptomycin resistance were sequenced for mutational analysis in the rpsL, rrs and gidB region. As the gidB protein has a complex substrate and no activity assay has yet been formulated, mutants of interest were subjected to in silico modelling and were structurally mapped together with active-site amino acid residues for assessment of the relevance to activity of the mutations found. RESULTS: Eight novel sense mutations and four novel mis-sense mutations in gidB were identified. Findings showed that active-site morphology is not only greatly affected by mutants lying in close proximity to the active-site pocket, but also by other mutations altering secondary-structure motifs and having an overall effect on protein structure. CONCLUSIONS: We conclude that gidB mutations address many unanswered questions and explain the whole story behind phenotypic streptomycin-resistant strains exhibiting no mutation in rpsL or rrs. They also validate the hypothesis of sequential progression of resistance from low to high due to the existence of gidB alterations in the genetic background.
OBJECTIVES: To evaluate gidB alterations for possible impact on the cumulative mechanism underlying the acquisition of high-level streptomycin resistance in Mycobacterium tuberculosis. METHODS: Fifty-two isolates with high streptomycin resistance and 23 isolates with low streptomycin resistance were sequenced for mutational analysis in the rpsL, rrs and gidB region. As the gidB protein has a complex substrate and no activity assay has yet been formulated, mutants of interest were subjected to in silico modelling and were structurally mapped together with active-site amino acid residues for assessment of the relevance to activity of the mutations found. RESULTS: Eight novel sense mutations and four novel mis-sense mutations in gidB were identified. Findings showed that active-site morphology is not only greatly affected by mutants lying in close proximity to the active-site pocket, but also by other mutations altering secondary-structure motifs and having an overall effect on protein structure. CONCLUSIONS: We conclude that gidB mutations address many unanswered questions and explain the whole story behind phenotypic streptomycin-resistant strains exhibiting no mutation in rpsL or rrs. They also validate the hypothesis of sequential progression of resistance from low to high due to the existence of gidB alterations in the genetic background.
Authors: Mohammad J Nasiri; Mehri Haeili; Mona Ghazi; Hossein Goudarzi; Ali Pormohammad; Abbas A Imani Fooladi; Mohammad M Feizabadi Journal: Front Microbiol Date: 2017-04-25 Impact factor: 5.640