Single molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes

DC FieldValueLanguage
dc.contributor.authorBarlag, Britta
dc.contributor.authorBeutel, Oliver
dc.contributor.authorJanning, Dennis
dc.contributor.authorCzarniak, Frederik
dc.contributor.authorRichter, Christian P.
dc.contributor.authorKommnick, Carina
dc.contributor.authorGoeser, Vera
dc.contributor.authorKurre, Rainer
dc.contributor.authorFabiani, Florian
dc.contributor.authorErhardt, Marc
dc.contributor.authorPiehler, Jacob
dc.contributor.authorHensel, Michael
dc.date.accessioned2021-12-23T16:12:49Z-
dc.date.available2021-12-23T16:12:49Z-
dc.date.issued2016
dc.identifier.issn20452322
dc.identifier.urihttps://osnascholar.ub.uni-osnabrueck.de/handle/unios/10290-
dc.description.abstractThe investigation of the subcellular localization, dynamics and interaction of proteins and protein complexes in prokaryotes is complicated by the small size of the cells. Super-resolution microscopy ( SRM) comprise various new techniques that allow light microscopy with a resolution that can be up to ten-fold higher than conventional light microscopy. Application of SRM techniques to living prokaryotes demands the introduction of suitable fluorescent probes, usually by fusion of proteins of interest to fluorescent proteins with properties compatible to SRM. Here we describe an approach that is based on the genetically encoded self-labelling enzymes HaloTag and SNAP-tag. Proteins of interest are fused to HaloTag or SNAP-tag and cell permeable substrates can be labelled with various SRM-compatible fluorochromes. Fusions of the enzyme tags to subunits of a type I secretion system ( T1SS), a T3SS, the flagellar rotor and a transcription factor were generated and analysed in living Salmonella enterica. The new approach is versatile in tagging proteins of interest in bacterial cells and allows to determine the number, relative subcellular localization and dynamics of protein complexes in living cells.
dc.description.sponsorshipHelmholtz AssociationHelmholtz Association [VH-NG-932]; European UnionEuropean Commission [334030]; President's Initiative of the Helmholtz Association of German Research Centers (HGF) [VH-GS-202]; Networking Funds of the Helmholtz Association of German Research Centers (HGF) [VH-GS-202]; DFGGerman Research Foundation (DFG)European Commission; Osnabruck University; This work was supported by the DFG by grants P4 (M.H.) and Z (M.H, J.P.) as part of SFB944 `Physiology and dynamics of cellular microcompartments'. We acknowledge support by the Helmholtz Association Young Investigator grant number VH-NG-932 and the People Program (Marie Curie Actions) of the European Unions' Seventh Framework Program grant number 334030 (M.E.). F.D.F. acknowledges support by the President's Initiative and Networking Funds of the Helmholtz Association of German Research Centers (HGF) under contract number VH-GS-202. support by the DFG and Open Access Publishing Fund of Osnabruck University. We thank Monika Nietschke and Nathalie Sander for technical and experimental support.
dc.language.isoen
dc.publisherNATURE PUBLISHING GROUP
dc.relation.ispartofSCIENTIFIC REPORTS
dc.subjectBACTERIAL-CELL BIOLOGY
dc.subjectCHEMOTAXIS
dc.subjectGENES
dc.subjectIII SECRETION
dc.subjectLOCALIZATION
dc.subjectMICROCOMPARTMENTS
dc.subjectMICROSCOPY
dc.subjectMultidisciplinary Sciences
dc.subjectNON-FIMBRIAL ADHESIN
dc.subjectORGANIZATION
dc.subjectScience & Technology - Other Topics
dc.subjectTRACKING
dc.titleSingle molecule super-resolution imaging of proteins in living Salmonella enterica using self-labelling enzymes
dc.typejournal article
dc.identifier.doi10.1038/srep31601
dc.identifier.isiISI:000381559000001
dc.description.volume6
dc.contributor.orcid0000-0001-6292-619X
dc.contributor.orcid0000-0001-6604-6253
dc.publisher.placeMACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND
dcterms.isPartOf.abbreviationSci Rep
dcterms.oaStatusGreen Published, Green Submitted, gold
crisitem.author.deptFB 05 - Biologie/Chemie-
crisitem.author.deptFB 05 - Biologie/Chemie-
crisitem.author.deptFB 05 - Biologie/Chemie-
crisitem.author.deptidfb05-
crisitem.author.deptidfb05-
crisitem.author.deptidfb05-
crisitem.author.orcid0000-0002-6872-6567-
crisitem.author.orcid0000-0002-2143-2270-
crisitem.author.orcid0000-0001-6604-6253-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.netidKuRa617-
crisitem.author.netidPiJa938-
crisitem.author.netidHeMi480-
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