Spontaneous Suppressors against Debilitating Transmembrane Mutants of <i>Ca</i>Mdr1 Disclose Novel Interdomain Communication via Signature Motifs of the Major Facilitator Superfamily
The Major Facilitator Superfamily (MFS) drug:H<sup>+</sup> antiporter <i>Ca</i>Mdr1, from <i>Candida albicans</i>, is responsible for the efflux of structurally diverse antifungals. MFS members share a common fold of 12–14 transmembrane helices (TMHs) forming two...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-05-01
|
| Series: | Journal of Fungi |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2309-608X/8/5/538 |
| _version_ | 1797498666251976704 |
|---|---|
| author | Suman Sharma Atanu Banerjee Alexis Moreno Archana Kumari Redhu Pierre Falson Rajendra Prasad |
| author_facet | Suman Sharma Atanu Banerjee Alexis Moreno Archana Kumari Redhu Pierre Falson Rajendra Prasad |
| author_sort | Suman Sharma |
| collection | DOAJ |
| description | The Major Facilitator Superfamily (MFS) drug:H<sup>+</sup> antiporter <i>Ca</i>Mdr1, from <i>Candida albicans</i>, is responsible for the efflux of structurally diverse antifungals. MFS members share a common fold of 12–14 transmembrane helices (TMHs) forming two N- and C-domains. Each domain is arranged in a pseudo-symmetric fold of two tandems of 3-TMHs that alternatively expose the drug-binding site towards the inside or the outside of the yeast to promote drug binding and release. MFS proteins show great diversity in primary structure and few conserved signature motifs, each thought to have a common function in the superfamily, although not yet clearly established. Here, we provide new information on these motifs by having screened a library of 64 drug transport-deficient mutants and their corresponding suppressors spontaneously addressing the deficiency. We found that five strains recovered the drug-resistance capacity by expressing <i>Ca</i>Mdr1 with a secondary mutation. The pairs of debilitating/rescuing residues are distributed either in the same TMH (T127A<sub>TMH1</sub>- > G140D<sub>TMH1</sub>) or 3-TMHs repeat (F216A<sub>TMH4</sub>- > G260A<sub>TMH5</sub>), at the hinge of 3-TMHs repeats tandems (R184A<sub>TMH3</sub>- > D235H<sub>TMH4</sub>, L480A<sub>TMH10</sub>- > A435T<sub>TMH9</sub>), and finally between the N- and C-domains (G230A<sub>TMH4</sub>- > P528H<sub>TMH12</sub>). Remarkably, most of these mutants belong to the different signature motifs, highlighting a mechanistic role and interplay thought to be conserved among MFS proteins. Results also point to the specific role of TMH11 in the interplay between the N- and C-domains in the inward- to outward-open conformational transition. |
| first_indexed | 2024-03-10T03:36:37Z |
| format | Article |
| id | doaj.art-547d9a98f5ea4f9887e427a4fa0fec77 |
| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-10T03:36:37Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-547d9a98f5ea4f9887e427a4fa0fec772023-11-23T11:42:33ZengMDPI AGJournal of Fungi2309-608X2022-05-018553810.3390/jof8050538Spontaneous Suppressors against Debilitating Transmembrane Mutants of <i>Ca</i>Mdr1 Disclose Novel Interdomain Communication via Signature Motifs of the Major Facilitator SuperfamilySuman Sharma0Atanu Banerjee1Alexis Moreno2Archana Kumari Redhu3Pierre Falson4Rajendra Prasad5Amity Institute of Biotechnology, Amity University Haryana, Gurugram 122413, IndiaAmity Institute of Biotechnology, Amity University Haryana, Gurugram 122413, IndiaDrug Resistance & Membrane Proteins Team, Molecular Microbiology and Structural Biochemistry Laboratory, Institut de Biologie et Chimie des Protéines, CNRS-Lyon 1 University Research lab n° 5086, 69367 Lyon, FranceACTREC, Tata Memorial Centre, Navi Mumbai 410210, IndiaDrug Resistance & Membrane Proteins Team, Molecular Microbiology and Structural Biochemistry Laboratory, Institut de Biologie et Chimie des Protéines, CNRS-Lyon 1 University Research lab n° 5086, 69367 Lyon, FranceAmity Institute of Biotechnology, Amity University Haryana, Gurugram 122413, IndiaThe Major Facilitator Superfamily (MFS) drug:H<sup>+</sup> antiporter <i>Ca</i>Mdr1, from <i>Candida albicans</i>, is responsible for the efflux of structurally diverse antifungals. MFS members share a common fold of 12–14 transmembrane helices (TMHs) forming two N- and C-domains. Each domain is arranged in a pseudo-symmetric fold of two tandems of 3-TMHs that alternatively expose the drug-binding site towards the inside or the outside of the yeast to promote drug binding and release. MFS proteins show great diversity in primary structure and few conserved signature motifs, each thought to have a common function in the superfamily, although not yet clearly established. Here, we provide new information on these motifs by having screened a library of 64 drug transport-deficient mutants and their corresponding suppressors spontaneously addressing the deficiency. We found that five strains recovered the drug-resistance capacity by expressing <i>Ca</i>Mdr1 with a secondary mutation. The pairs of debilitating/rescuing residues are distributed either in the same TMH (T127A<sub>TMH1</sub>- > G140D<sub>TMH1</sub>) or 3-TMHs repeat (F216A<sub>TMH4</sub>- > G260A<sub>TMH5</sub>), at the hinge of 3-TMHs repeats tandems (R184A<sub>TMH3</sub>- > D235H<sub>TMH4</sub>, L480A<sub>TMH10</sub>- > A435T<sub>TMH9</sub>), and finally between the N- and C-domains (G230A<sub>TMH4</sub>- > P528H<sub>TMH12</sub>). Remarkably, most of these mutants belong to the different signature motifs, highlighting a mechanistic role and interplay thought to be conserved among MFS proteins. Results also point to the specific role of TMH11 in the interplay between the N- and C-domains in the inward- to outward-open conformational transition.https://www.mdpi.com/2309-608X/8/5/538<i>Candida albicans</i>antifungal resistanceMFS transporter<i>Ca</i>Mdr1drug:H<sup>+</sup> antiporter 1interdomain crosstalk |
| spellingShingle | Suman Sharma Atanu Banerjee Alexis Moreno Archana Kumari Redhu Pierre Falson Rajendra Prasad Spontaneous Suppressors against Debilitating Transmembrane Mutants of <i>Ca</i>Mdr1 Disclose Novel Interdomain Communication via Signature Motifs of the Major Facilitator Superfamily Journal of Fungi <i>Candida albicans</i> antifungal resistance MFS transporter <i>Ca</i>Mdr1 drug:H<sup>+</sup> antiporter 1 interdomain crosstalk |
| title | Spontaneous Suppressors against Debilitating Transmembrane Mutants of <i>Ca</i>Mdr1 Disclose Novel Interdomain Communication via Signature Motifs of the Major Facilitator Superfamily |
| title_full | Spontaneous Suppressors against Debilitating Transmembrane Mutants of <i>Ca</i>Mdr1 Disclose Novel Interdomain Communication via Signature Motifs of the Major Facilitator Superfamily |
| title_fullStr | Spontaneous Suppressors against Debilitating Transmembrane Mutants of <i>Ca</i>Mdr1 Disclose Novel Interdomain Communication via Signature Motifs of the Major Facilitator Superfamily |
| title_full_unstemmed | Spontaneous Suppressors against Debilitating Transmembrane Mutants of <i>Ca</i>Mdr1 Disclose Novel Interdomain Communication via Signature Motifs of the Major Facilitator Superfamily |
| title_short | Spontaneous Suppressors against Debilitating Transmembrane Mutants of <i>Ca</i>Mdr1 Disclose Novel Interdomain Communication via Signature Motifs of the Major Facilitator Superfamily |
| title_sort | spontaneous suppressors against debilitating transmembrane mutants of i ca i mdr1 disclose novel interdomain communication via signature motifs of the major facilitator superfamily |
| topic | <i>Candida albicans</i> antifungal resistance MFS transporter <i>Ca</i>Mdr1 drug:H<sup>+</sup> antiporter 1 interdomain crosstalk |
| url | https://www.mdpi.com/2309-608X/8/5/538 |
| work_keys_str_mv | AT sumansharma spontaneoussuppressorsagainstdebilitatingtransmembranemutantsoficaimdr1disclosenovelinterdomaincommunicationviasignaturemotifsofthemajorfacilitatorsuperfamily AT atanubanerjee spontaneoussuppressorsagainstdebilitatingtransmembranemutantsoficaimdr1disclosenovelinterdomaincommunicationviasignaturemotifsofthemajorfacilitatorsuperfamily AT alexismoreno spontaneoussuppressorsagainstdebilitatingtransmembranemutantsoficaimdr1disclosenovelinterdomaincommunicationviasignaturemotifsofthemajorfacilitatorsuperfamily AT archanakumariredhu spontaneoussuppressorsagainstdebilitatingtransmembranemutantsoficaimdr1disclosenovelinterdomaincommunicationviasignaturemotifsofthemajorfacilitatorsuperfamily AT pierrefalson spontaneoussuppressorsagainstdebilitatingtransmembranemutantsoficaimdr1disclosenovelinterdomaincommunicationviasignaturemotifsofthemajorfacilitatorsuperfamily AT rajendraprasad spontaneoussuppressorsagainstdebilitatingtransmembranemutantsoficaimdr1disclosenovelinterdomaincommunicationviasignaturemotifsofthemajorfacilitatorsuperfamily |