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Biochemical and phenotypic characterisation of the Mycobacterium smegmatis transporter UspABC
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Karlikowska, Magdalena, Singh, Albel, Bhatt, Apoorva, Ott, Sascha, Bottrill, Andrew R., Besra, Gurdyal S. and Fullam, Elizabeth (2021) Biochemical and phenotypic characterisation of the Mycobacterium smegmatis transporter UspABC. The Cell Surface, 7 . 100052. doi:10.1016/j.tcsw.2021.100052
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Official URL: http://dx.doi.org/10.1016/j.tcsw.2021.100052
Abstract
Mycobacterium tuberculosis (Mtb) is an intracellular human pathogen that has evolved to survive in a nutrient limited environment within the host for decades. Accordingly, Mtb has developed strategies to acquire scarce nutrients and the mycobacterial transporter systems provide an important route for the import of key energy sources. However, the physiological role of the Mtb transporters and their substrate preference(s) are poorly characterised. Previous studies have established that the Mtb UspC solute-binding domain recognises amino- and phosphorylated-sugars, indicating that the mycobacterial UspABC transporter plays a key role in the import of peptidoglycan precursors. Herein, we have used a wide array of approaches to investigate the role of UspABC in Mycobacterium smegmatis by analysis of mutant strains that either lack the solute binding domain: ΔuspC or the entire transport complex: ΔuspABC. Analysis of mycobacterial transcripts shows that the uspABC system is functionally expressed in mycobacteria as a contiguous reading frame. Topology mapping confirms an Nin-Cin orientation of the UspAB integral membrane spanning domains. Phenotypic microarray profiling of commercially available sugars suggests, unexpectedly, that the uspC and ΔuspABC mutants had different carbon utilisation profiles and that neither strain utilised glucose-1-phosphate. Furthermore, proteomics analysis showed an alteration in the abundance of proteins involved in sugar and lipid metabolism, crucial for cell envelope synthesis, and we propose that UspABC has an important role in determining the interplay between these pathways.
| Item Type: | Journal Article | |||||||||||||||||||||||||||||||||
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| Subjects: | Q Science > QP Physiology Q Science > QR Microbiology |
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| Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School |
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| Library of Congress Subject Headings (LCSH): | Mycobacteria , Mycobacterium tuberculosis, Tuberculosis -- Molecular aspects, Host-virus relationships, ATP-binding cassette transporters, Carrier proteins | |||||||||||||||||||||||||||||||||
| Journal or Publication Title: | The Cell Surface | |||||||||||||||||||||||||||||||||
| Publisher: | Elsevier | |||||||||||||||||||||||||||||||||
| ISSN: | 2468-2330 | |||||||||||||||||||||||||||||||||
| Official Date: | December 2021 | |||||||||||||||||||||||||||||||||
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| Volume: | 7 | |||||||||||||||||||||||||||||||||
| Article Number: | 100052 | |||||||||||||||||||||||||||||||||
| DOI: | 10.1016/j.tcsw.2021.100052 | |||||||||||||||||||||||||||||||||
| Status: | Peer Reviewed | |||||||||||||||||||||||||||||||||
| Publication Status: | Published | |||||||||||||||||||||||||||||||||
| Access rights to Published version: | Open Access | |||||||||||||||||||||||||||||||||
| RIOXX Funder/Project Grant: |
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