Translation Stress Positively Regulates MscL-Dependent Excretion of Cytoplasmic Proteins

ABSTRACT The apparent mislocalization or excretion of cytoplasmic proteins is a commonly observed phenomenon in both bacteria and eukaryotes. However, reports on the mechanistic basis and the cellular function of this so-called “nonclassical protein secretion” are limited. Here we report that protei...

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Autores principales:	Rosa Morra, Francesco Del Carratore, Howbeer Muhamadali, Luminita Gabriela Horga, Samantha Halliwell, Royston Goodacre, Rainer Breitling, Neil Dixon
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	ArfA MscL osmotic stress protein excretion translation stress Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/5ed4c5819e2548358622052199e81c80
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spelling	oai:doaj.org-article:5ed4c5819e2548358622052199e81c802021-11-15T15:53:26ZTranslation Stress Positively Regulates MscL-Dependent Excretion of Cytoplasmic Proteins10.1128/mBio.02118-172150-7511https://doaj.org/article/5ed4c5819e2548358622052199e81c802018-03-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02118-17https://doaj.org/toc/2150-7511ABSTRACT The apparent mislocalization or excretion of cytoplasmic proteins is a commonly observed phenomenon in both bacteria and eukaryotes. However, reports on the mechanistic basis and the cellular function of this so-called “nonclassical protein secretion” are limited. Here we report that protein overexpression in recombinant cells and antibiotic-induced translation stress in wild-type Escherichia coli cells both lead to excretion of cytoplasmic protein (ECP). Condition-specific metabolomic and proteomic analyses, combined with genetic knockouts, indicate a role for both the large mechanosensitive channel (MscL) and the alternative ribosome rescue factor A (ArfA) in ECP. Collectively, the findings indicate that MscL-dependent protein excretion is positively regulated in response to both osmotic stress and arfA-mediated translational stress. IMPORTANCE Protein translocation is an essential feature of cellular organisms. Bacteria, like all single-cell organisms, interact with their environment by translocation of proteins across their cell membranes via dedicated secretion pathways. Proteins destined for secretion are directed toward the secretion pathways by the presence of specific signal peptides. This study demonstrates that under conditions of both osmotic stress and translation stress, E. coli cells undergo an excretion phenomenon whereby signal peptide-less proteins are translocated across both the inner and outer cell membranes into the extracellular environment. Confirming the presence of alternative translocation/excretion pathways and understanding their function and regulation are thus important for fundamental microbiology and biotechnology.Rosa MorraFrancesco Del CarratoreHowbeer MuhamadaliLuminita Gabriela HorgaSamantha HalliwellRoyston GoodacreRainer BreitlingNeil DixonAmerican Society for MicrobiologyarticleArfAMscLosmotic stressprotein excretiontranslation stressMicrobiologyQR1-502ENmBio, Vol 9, Iss 1 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	ArfA MscL osmotic stress protein excretion translation stress Microbiology QR1-502
spellingShingle	ArfA MscL osmotic stress protein excretion translation stress Microbiology QR1-502 Rosa Morra Francesco Del Carratore Howbeer Muhamadali Luminita Gabriela Horga Samantha Halliwell Royston Goodacre Rainer Breitling Neil Dixon Translation Stress Positively Regulates MscL-Dependent Excretion of Cytoplasmic Proteins
description	ABSTRACT The apparent mislocalization or excretion of cytoplasmic proteins is a commonly observed phenomenon in both bacteria and eukaryotes. However, reports on the mechanistic basis and the cellular function of this so-called “nonclassical protein secretion” are limited. Here we report that protein overexpression in recombinant cells and antibiotic-induced translation stress in wild-type Escherichia coli cells both lead to excretion of cytoplasmic protein (ECP). Condition-specific metabolomic and proteomic analyses, combined with genetic knockouts, indicate a role for both the large mechanosensitive channel (MscL) and the alternative ribosome rescue factor A (ArfA) in ECP. Collectively, the findings indicate that MscL-dependent protein excretion is positively regulated in response to both osmotic stress and arfA-mediated translational stress. IMPORTANCE Protein translocation is an essential feature of cellular organisms. Bacteria, like all single-cell organisms, interact with their environment by translocation of proteins across their cell membranes via dedicated secretion pathways. Proteins destined for secretion are directed toward the secretion pathways by the presence of specific signal peptides. This study demonstrates that under conditions of both osmotic stress and translation stress, E. coli cells undergo an excretion phenomenon whereby signal peptide-less proteins are translocated across both the inner and outer cell membranes into the extracellular environment. Confirming the presence of alternative translocation/excretion pathways and understanding their function and regulation are thus important for fundamental microbiology and biotechnology.
format	article
author	Rosa Morra Francesco Del Carratore Howbeer Muhamadali Luminita Gabriela Horga Samantha Halliwell Royston Goodacre Rainer Breitling Neil Dixon
author_facet	Rosa Morra Francesco Del Carratore Howbeer Muhamadali Luminita Gabriela Horga Samantha Halliwell Royston Goodacre Rainer Breitling Neil Dixon
author_sort	Rosa Morra
title	Translation Stress Positively Regulates MscL-Dependent Excretion of Cytoplasmic Proteins
title_short	Translation Stress Positively Regulates MscL-Dependent Excretion of Cytoplasmic Proteins
title_full	Translation Stress Positively Regulates MscL-Dependent Excretion of Cytoplasmic Proteins
title_fullStr	Translation Stress Positively Regulates MscL-Dependent Excretion of Cytoplasmic Proteins
title_full_unstemmed	Translation Stress Positively Regulates MscL-Dependent Excretion of Cytoplasmic Proteins
title_sort	translation stress positively regulates mscl-dependent excretion of cytoplasmic proteins
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/5ed4c5819e2548358622052199e81c80
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Translation Stress Positively Regulates MscL-Dependent Excretion of Cytoplasmic Proteins

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