The extracellular loop of Man-PTS subunit IID is responsible for the sensitivity of Lactococcus garvieae to garvicins A, B and C

Abstract Mannose phosphotransferase system (Man-PTS) serves as a receptor for several bacteriocins in sensitive bacterial cells, namely subclass IIa bacteriocins (pediocin-like; pediocins) and subclass IId ones - lactococcin A (LcnA), lactococcin B (LcnB) and garvicin Q (GarQ). Here, to identify the...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Aleksandra Tymoszewska, Dzung B. Diep, Tamara Aleksandrzak-Piekarczyk
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Mannose Phosphotransferase System (Man-PTS) Lactococcin Pediocin Lactis Garvieae Strains Medicine R Science Q
Acceso en línea:	https://doaj.org/article/febdb3c50100445abf5626ee72c4a220
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:febdb3c50100445abf5626ee72c4a220
record_format	dspace
spelling	oai:doaj.org-article:febdb3c50100445abf5626ee72c4a2202021-12-02T15:07:52ZThe extracellular loop of Man-PTS subunit IID is responsible for the sensitivity of Lactococcus garvieae to garvicins A, B and C10.1038/s41598-018-34087-22045-2322https://doaj.org/article/febdb3c50100445abf5626ee72c4a2202018-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-34087-2https://doaj.org/toc/2045-2322Abstract Mannose phosphotransferase system (Man-PTS) serves as a receptor for several bacteriocins in sensitive bacterial cells, namely subclass IIa bacteriocins (pediocin-like; pediocins) and subclass IId ones - lactococcin A (LcnA), lactococcin B (LcnB) and garvicin Q (GarQ). Here, to identify the receptor for three other narrow-spectrum subclass IId bacteriocins - garvicins A, B and C (GarA-C) Lactococcus garvieae mutants resistant to bacteriocins were generated and sequenced to look for mutations responsible for resistance. Spontaneous mutants had their whole genome sequenced while in mutants obtained by integration of pGhost9::ISS1 regions flanking the integration site were sequenced. For both types of mutants mutations were found in genes encoding Man-PTS components IIC and IID indicating that Man-PTS likely serves as the receptor for these bacteriocins as well. This was subsequently confirmed by deletion of the man-PTS operon in the bacteriocin-sensitive L. garvieae IBB3403, which resulted in resistant cells, and by heterologous expression of appropriate man-PTS genes in the resistant Lactococcus lactis strains, which resulted in sensitive cells. GarA, GarB, GarC and other Man-PTS-targeting bacteriocins differ in the amino acid sequence and activity spectrum, suggesting that they interact with the receptor through distinct binding patterns. Comparative analyses and genetic studies identified a previously unrecognized extracellular loop of Man-PTS subunit IID (γ+) implicated in the L. garvieae sensitivity to the bacteriocins studied here. Additionally, individual amino acids localized mostly in the sugar channel-forming transmembrane parts of subunit IIC or in the extracellular parts of IID likely involved in the interaction with each bacteriocin were specified. Finally, template-based 3D models of Man-PTS subunits IIC and IID were built to allow a deeper insight into the Man-PTS structure and functioning.Aleksandra TymoszewskaDzung B. DiepTamara Aleksandrzak-PiekarczykNature PortfolioarticleMannose Phosphotransferase System (Man-PTS)LactococcinPediocinLactisGarvieae StrainsMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-15 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Mannose Phosphotransferase System (Man-PTS) Lactococcin Pediocin Lactis Garvieae Strains Medicine R Science Q
spellingShingle	Mannose Phosphotransferase System (Man-PTS) Lactococcin Pediocin Lactis Garvieae Strains Medicine R Science Q Aleksandra Tymoszewska Dzung B. Diep Tamara Aleksandrzak-Piekarczyk The extracellular loop of Man-PTS subunit IID is responsible for the sensitivity of Lactococcus garvieae to garvicins A, B and C
description	Abstract Mannose phosphotransferase system (Man-PTS) serves as a receptor for several bacteriocins in sensitive bacterial cells, namely subclass IIa bacteriocins (pediocin-like; pediocins) and subclass IId ones - lactococcin A (LcnA), lactococcin B (LcnB) and garvicin Q (GarQ). Here, to identify the receptor for three other narrow-spectrum subclass IId bacteriocins - garvicins A, B and C (GarA-C) Lactococcus garvieae mutants resistant to bacteriocins were generated and sequenced to look for mutations responsible for resistance. Spontaneous mutants had their whole genome sequenced while in mutants obtained by integration of pGhost9::ISS1 regions flanking the integration site were sequenced. For both types of mutants mutations were found in genes encoding Man-PTS components IIC and IID indicating that Man-PTS likely serves as the receptor for these bacteriocins as well. This was subsequently confirmed by deletion of the man-PTS operon in the bacteriocin-sensitive L. garvieae IBB3403, which resulted in resistant cells, and by heterologous expression of appropriate man-PTS genes in the resistant Lactococcus lactis strains, which resulted in sensitive cells. GarA, GarB, GarC and other Man-PTS-targeting bacteriocins differ in the amino acid sequence and activity spectrum, suggesting that they interact with the receptor through distinct binding patterns. Comparative analyses and genetic studies identified a previously unrecognized extracellular loop of Man-PTS subunit IID (γ+) implicated in the L. garvieae sensitivity to the bacteriocins studied here. Additionally, individual amino acids localized mostly in the sugar channel-forming transmembrane parts of subunit IIC or in the extracellular parts of IID likely involved in the interaction with each bacteriocin were specified. Finally, template-based 3D models of Man-PTS subunits IIC and IID were built to allow a deeper insight into the Man-PTS structure and functioning.
format	article
author	Aleksandra Tymoszewska Dzung B. Diep Tamara Aleksandrzak-Piekarczyk
author_facet	Aleksandra Tymoszewska Dzung B. Diep Tamara Aleksandrzak-Piekarczyk
author_sort	Aleksandra Tymoszewska
title	The extracellular loop of Man-PTS subunit IID is responsible for the sensitivity of Lactococcus garvieae to garvicins A, B and C
title_short	The extracellular loop of Man-PTS subunit IID is responsible for the sensitivity of Lactococcus garvieae to garvicins A, B and C
title_full	The extracellular loop of Man-PTS subunit IID is responsible for the sensitivity of Lactococcus garvieae to garvicins A, B and C
title_fullStr	The extracellular loop of Man-PTS subunit IID is responsible for the sensitivity of Lactococcus garvieae to garvicins A, B and C
title_full_unstemmed	The extracellular loop of Man-PTS subunit IID is responsible for the sensitivity of Lactococcus garvieae to garvicins A, B and C
title_sort	extracellular loop of man-pts subunit iid is responsible for the sensitivity of lactococcus garvieae to garvicins a, b and c
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/febdb3c50100445abf5626ee72c4a220
work_keys_str_mv	AT aleksandratymoszewska theextracellularloopofmanptssubunitiidisresponsibleforthesensitivityoflactococcusgarvieaetogarvicinsabandc AT dzungbdiep theextracellularloopofmanptssubunitiidisresponsibleforthesensitivityoflactococcusgarvieaetogarvicinsabandc AT tamaraaleksandrzakpiekarczyk theextracellularloopofmanptssubunitiidisresponsibleforthesensitivityoflactococcusgarvieaetogarvicinsabandc AT aleksandratymoszewska extracellularloopofmanptssubunitiidisresponsibleforthesensitivityoflactococcusgarvieaetogarvicinsabandc AT dzungbdiep extracellularloopofmanptssubunitiidisresponsibleforthesensitivityoflactococcusgarvieaetogarvicinsabandc AT tamaraaleksandrzakpiekarczyk extracellularloopofmanptssubunitiidisresponsibleforthesensitivityoflactococcusgarvieaetogarvicinsabandc
_version_	1718388427470143488

The extracellular loop of Man-PTS subunit IID is responsible for the sensitivity of Lactococcus garvieae to garvicins A, B and C

Ejemplares similares