title: Fluorescence-based sensors to monitor localization and functions of linear and K63-linked ubiquitin chains in cells.
abstract: Ubiquitin chains modify a major subset of the proteome, but detection of ubiquitin signaling dynamics and localization is limited due to a lack of appropriate tools. Here, we employ ubiquitin-binding domain (UBD)-based fluorescent sensors to monitor linear and K63-linked chains in vitro and in vivo. We utilize the UBD in NEMO and ABIN (UBAN) for detection of linear chains, and RAP80 ubiquitin-interacting motif (UIM) and TAB2 Npl4 zinc finger (NZF) domains to detect K63 chains. Linear and K63 sensors decorated the ubiquitin coat surrounding cytosolic Salmonella during bacterial autophagy, whereas K63 sensors selectively monitored Parkin-induced mitophagy and DNA damage responses in fixed and living cells. In addition, linear and K63 sensors could be used to monitor endogenous signaling pathways, as demonstrated by their ability to differentially interfere with TNF- and IL-1-induced NF-κB pathway. We propose that UBD-based biosensors could serve as prototypes to track and trace other chain types and ubiquitin-like signals in vivo.
authors: Institute of Biochemistry II, Goethe University School of Medicine, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany.
authors: Sjoerd J L
authors: SJ
authors: van Wijk
authors: Evgenij
authors: E
authors: Fiskin
authors: Mateusz
authors: M
authors: Putyrski
authors: Francesco
authors: F
authors: Pampaloni
authors: Jian
authors: J
authors: Hou
authors: Philipp
authors: P
authors: Wild
authors: Tobias
authors: T
authors: Kensche
authors: Hernan E
authors: HE
authors: Grecco
authors: Philippe
authors: P
authors: Bastiaens
authors: Ivan
authors: I
authors: Dikic
mesh_terms: D015374:Biosensing Techniques; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012480:Salmonella Infections / Q000378:metabolism; D015398:Signal Transduction; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D016328:NF-kappa B; D011994:Recombinant Proteins; D025801:Ubiquitin; D049452:Green Fluorescent Proteins