An Intracellular Nanotrap Redirects Proteins and Organelles in Live Bacteria

ABSTRACT Owing to their small size and enhanced stability, nanobodies derived from camelids have previously been used for the construction of intracellular “nanotraps,” which enable redirection and manipulation of green fluorescent protein (GFP)-tagged targets within living plant and animal cells....

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Autores principales:	Sarah Borg, Felix Popp, Julia Hofmann, Heinrich Leonhardt, Ulrich Rothbauer, Dirk Schüler
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2015
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/bb87def48bd1496cbe7ffe22d0649c08
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spelling	oai:doaj.org-article:bb87def48bd1496cbe7ffe22d0649c082021-11-15T15:41:19ZAn Intracellular Nanotrap Redirects Proteins and Organelles in Live Bacteria10.1128/mBio.02117-142150-7511https://doaj.org/article/bb87def48bd1496cbe7ffe22d0649c082015-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02117-14https://doaj.org/toc/2150-7511ABSTRACT Owing to their small size and enhanced stability, nanobodies derived from camelids have previously been used for the construction of intracellular “nanotraps,” which enable redirection and manipulation of green fluorescent protein (GFP)-tagged targets within living plant and animal cells. By taking advantage of intracellular compartmentalization in the magnetic bacterium Magnetospirillum gryphiswaldense, we demonstrate that proteins and even entire organelles can be retargeted also within prokaryotic cells by versatile nanotrap technology. Expression of multivalent GFP-binding nanobodies on magnetosomes ectopically recruited the chemotaxis protein CheW1-GFP from polar chemoreceptor clusters to the midcell, resulting in a gradual knockdown of aerotaxis. Conversely, entire magnetosome chains could be redirected from the midcell and tethered to one of the cell poles. Similar approaches could potentially be used for building synthetic cellular structures and targeted protein knockdowns in other bacteria. IMPORTANCE Intrabodies are commonly used in eukaryotic systems for intracellular analysis and manipulation of proteins within distinct subcellular compartments. In particular, so-called nanobodies have great potential for synthetic biology approaches because they can be expressed easily in heterologous hosts and actively interact with intracellular targets, for instance, by the construction of intracellular “nanotraps” in living animal and plant cells. Although prokaryotic cells also exhibit a considerable degree of intracellular organization, there are few tools available equivalent to the well-established methods used in eukaryotes. Here, we demonstrate the ectopic retargeting and depletion of polar membrane proteins and entire organelles to distinct compartments in a magnetotactic bacterium, resulting in a gradual knockdown of magneto-aerotaxis. This intracellular nanotrap approach has the potential to be applied in other bacteria for building synthetic cellular structures, manipulating protein function, and creating gradual targeted knockdowns. Our findings provide a proof of principle for the universal use of fluorescently tagged proteins as targets for nanotraps to fulfill these tasks.Sarah BorgFelix PoppJulia HofmannHeinrich LeonhardtUlrich RothbauerDirk SchülerAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 6, Iss 1 (2015)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Sarah Borg Felix Popp Julia Hofmann Heinrich Leonhardt Ulrich Rothbauer Dirk Schüler An Intracellular Nanotrap Redirects Proteins and Organelles in Live Bacteria
description	ABSTRACT Owing to their small size and enhanced stability, nanobodies derived from camelids have previously been used for the construction of intracellular “nanotraps,” which enable redirection and manipulation of green fluorescent protein (GFP)-tagged targets within living plant and animal cells. By taking advantage of intracellular compartmentalization in the magnetic bacterium Magnetospirillum gryphiswaldense, we demonstrate that proteins and even entire organelles can be retargeted also within prokaryotic cells by versatile nanotrap technology. Expression of multivalent GFP-binding nanobodies on magnetosomes ectopically recruited the chemotaxis protein CheW1-GFP from polar chemoreceptor clusters to the midcell, resulting in a gradual knockdown of aerotaxis. Conversely, entire magnetosome chains could be redirected from the midcell and tethered to one of the cell poles. Similar approaches could potentially be used for building synthetic cellular structures and targeted protein knockdowns in other bacteria. IMPORTANCE Intrabodies are commonly used in eukaryotic systems for intracellular analysis and manipulation of proteins within distinct subcellular compartments. In particular, so-called nanobodies have great potential for synthetic biology approaches because they can be expressed easily in heterologous hosts and actively interact with intracellular targets, for instance, by the construction of intracellular “nanotraps” in living animal and plant cells. Although prokaryotic cells also exhibit a considerable degree of intracellular organization, there are few tools available equivalent to the well-established methods used in eukaryotes. Here, we demonstrate the ectopic retargeting and depletion of polar membrane proteins and entire organelles to distinct compartments in a magnetotactic bacterium, resulting in a gradual knockdown of magneto-aerotaxis. This intracellular nanotrap approach has the potential to be applied in other bacteria for building synthetic cellular structures, manipulating protein function, and creating gradual targeted knockdowns. Our findings provide a proof of principle for the universal use of fluorescently tagged proteins as targets for nanotraps to fulfill these tasks.
format	article
author	Sarah Borg Felix Popp Julia Hofmann Heinrich Leonhardt Ulrich Rothbauer Dirk Schüler
author_facet	Sarah Borg Felix Popp Julia Hofmann Heinrich Leonhardt Ulrich Rothbauer Dirk Schüler
author_sort	Sarah Borg
title	An Intracellular Nanotrap Redirects Proteins and Organelles in Live Bacteria
title_short	An Intracellular Nanotrap Redirects Proteins and Organelles in Live Bacteria
title_full	An Intracellular Nanotrap Redirects Proteins and Organelles in Live Bacteria
title_fullStr	An Intracellular Nanotrap Redirects Proteins and Organelles in Live Bacteria
title_full_unstemmed	An Intracellular Nanotrap Redirects Proteins and Organelles in Live Bacteria
title_sort	intracellular nanotrap redirects proteins and organelles in live bacteria
publisher	American Society for Microbiology
publishDate	2015
url	https://doaj.org/article/bb87def48bd1496cbe7ffe22d0649c08
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An Intracellular Nanotrap Redirects Proteins and Organelles in Live Bacteria

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