A Loss of Function in LprG−Rv1410c Homologues Attenuates Growth during Biofilm Formation in <i>Mycobacterium smegmatis</i>
MmpL (mycobacterial membrane protein large) proteins are integral membrane proteins that have been implicated in the biosynthesis and/or transport of mycobacterial cell wall lipids. Given the cellular location of these proteins, however, it is unclear how cell wall lipids are transported beyond the...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-11-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/12/12/1375 |
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| author | Lisa-Marie Nisbett Mary L. Previti Jessica C. Seeliger |
| author_facet | Lisa-Marie Nisbett Mary L. Previti Jessica C. Seeliger |
| author_sort | Lisa-Marie Nisbett |
| collection | DOAJ |
| description | MmpL (mycobacterial membrane protein large) proteins are integral membrane proteins that have been implicated in the biosynthesis and/or transport of mycobacterial cell wall lipids. Given the cellular location of these proteins, however, it is unclear how cell wall lipids are transported beyond the inner membrane. Moreover, given that mycobacteria grow at the poles, we also do not understand how new cell wall is added in a highly localized and presumably coordinated manner. Here, we examine the relationship between two lipid transport pathways associated with the proteins MmpL11 and LprG−Rv1410c. The lipoprotein LprG has been shown to interact with proteins involved in cell wall processes including MmpL11, which is required in biofilms for the surface localization of certain lipids. Here we report that deletion of <i>mmpL11</i> (<i>MSMEG_0241</i>) or the <i>lprG−rv1410c</i> operon homologues <i>MSMEG_3070−3069</i> in <i>Mycobacterium smegmatis</i> produced similar biofilm defects that were distinct from that of the previously reported <i>mmpL11</i> transposon insertion mutant. Analysis of pellicle biofilms, bacterial growth, lipid profiles, and gene expression revealed that the biofilm phenotypes could not be directly explained by changes in the synthesis or localization of biofilm-related lipids or the expression of biofilm-related genes. Instead, the shared biofilm phenotype between Δ<i>MSMEG_3070−3069</i> and Δ<i>mmpL11</i> may be related to their modest growth defect, while the origins of the distinct <i>mmpL11::Tn</i> biofilm defect remain unclear. Our findings suggest that the mechanisms that drive pellicle biofilm formation in <i>M. smegmatis</i> are not connected to crosstalk between the LprG−Rv1410c and MmpL11 pathways and that any functional interaction between these proteins does not relate directly to their lipid transport function. |
| first_indexed | 2024-03-08T20:27:58Z |
| format | Article |
| id | doaj.art-5d21669735104cf2b757fbf41d35ea7d |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-08T20:27:58Z |
| publishDate | 2023-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-5d21669735104cf2b757fbf41d35ea7d2023-12-22T14:30:44ZengMDPI AGPathogens2076-08172023-11-011212137510.3390/pathogens12121375A Loss of Function in LprG−Rv1410c Homologues Attenuates Growth during Biofilm Formation in <i>Mycobacterium smegmatis</i>Lisa-Marie Nisbett0Mary L. Previti1Jessica C. Seeliger2Department of Pharmacological Sciences, Stony Brook University, 100 Nicolls Road, Stony Brook, NY 11794, USADepartment of Pharmacological Sciences, Stony Brook University, 100 Nicolls Road, Stony Brook, NY 11794, USADepartment of Pharmacological Sciences, Stony Brook University, 100 Nicolls Road, Stony Brook, NY 11794, USAMmpL (mycobacterial membrane protein large) proteins are integral membrane proteins that have been implicated in the biosynthesis and/or transport of mycobacterial cell wall lipids. Given the cellular location of these proteins, however, it is unclear how cell wall lipids are transported beyond the inner membrane. Moreover, given that mycobacteria grow at the poles, we also do not understand how new cell wall is added in a highly localized and presumably coordinated manner. Here, we examine the relationship between two lipid transport pathways associated with the proteins MmpL11 and LprG−Rv1410c. The lipoprotein LprG has been shown to interact with proteins involved in cell wall processes including MmpL11, which is required in biofilms for the surface localization of certain lipids. Here we report that deletion of <i>mmpL11</i> (<i>MSMEG_0241</i>) or the <i>lprG−rv1410c</i> operon homologues <i>MSMEG_3070−3069</i> in <i>Mycobacterium smegmatis</i> produced similar biofilm defects that were distinct from that of the previously reported <i>mmpL11</i> transposon insertion mutant. Analysis of pellicle biofilms, bacterial growth, lipid profiles, and gene expression revealed that the biofilm phenotypes could not be directly explained by changes in the synthesis or localization of biofilm-related lipids or the expression of biofilm-related genes. Instead, the shared biofilm phenotype between Δ<i>MSMEG_3070−3069</i> and Δ<i>mmpL11</i> may be related to their modest growth defect, while the origins of the distinct <i>mmpL11::Tn</i> biofilm defect remain unclear. Our findings suggest that the mechanisms that drive pellicle biofilm formation in <i>M. smegmatis</i> are not connected to crosstalk between the LprG−Rv1410c and MmpL11 pathways and that any functional interaction between these proteins does not relate directly to their lipid transport function.https://www.mdpi.com/2076-0817/12/12/1375mycobacteriacell wallbiofilmslipid transportMmpL11LprG |
| spellingShingle | Lisa-Marie Nisbett Mary L. Previti Jessica C. Seeliger A Loss of Function in LprG−Rv1410c Homologues Attenuates Growth during Biofilm Formation in <i>Mycobacterium smegmatis</i> Pathogens mycobacteria cell wall biofilms lipid transport MmpL11 LprG |
| title | A Loss of Function in LprG−Rv1410c Homologues Attenuates Growth during Biofilm Formation in <i>Mycobacterium smegmatis</i> |
| title_full | A Loss of Function in LprG−Rv1410c Homologues Attenuates Growth during Biofilm Formation in <i>Mycobacterium smegmatis</i> |
| title_fullStr | A Loss of Function in LprG−Rv1410c Homologues Attenuates Growth during Biofilm Formation in <i>Mycobacterium smegmatis</i> |
| title_full_unstemmed | A Loss of Function in LprG−Rv1410c Homologues Attenuates Growth during Biofilm Formation in <i>Mycobacterium smegmatis</i> |
| title_short | A Loss of Function in LprG−Rv1410c Homologues Attenuates Growth during Biofilm Formation in <i>Mycobacterium smegmatis</i> |
| title_sort | loss of function in lprg rv1410c homologues attenuates growth during biofilm formation in i mycobacterium smegmatis i |
| topic | mycobacteria cell wall biofilms lipid transport MmpL11 LprG |
| url | https://www.mdpi.com/2076-0817/12/12/1375 |
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