![]() ![]() ![]() Temperature-dependence (PMID:28115706) reversibility of formation and dissolution (PMID:28115706)ġ) rDNA (modulates the kinetics, variability of the process but not required) Probable ATP-dependent RNA helicase pitchoune Disease-associated SPOP-mutations that lead to the accumulation of proto-oncogenic proteins interfere with phase separation and co-localization in membraneless organelles, suggesting that substrate-directed phase separation of this E3 ligase underlies the regulation of ubiquitin-dependent proteostasis (PMID:30244836). Enzymatic activity correlates with cellular co-localization and in vitro mesoscale assembly formation. Substrates include the death-domain-associated protein (DAXX), androgen receptor (AR), and other important signaling cascade effectors, epigenetic modifiers and hormone signaling effectors, these contain multiple SPOP-binding (SB) motifs in their IDRs. Substrates trigger phase separation of SPOP in vitro and co-localization in membraneless organelles in cells. SPOP is a substrate adaptor of the cullin3-RING ubiquitin ligase and localizes to nuclear speckles. Mutations in the tumor suppressor SPOP (speckle-type POZ protein) cause prostate, breast and other solid tumors. Nuclear body nuclear protein granule SPOP/DAXX body Multivalent domain-motif interactions (PMID:30244836) linear oligomerization/self-association (PMID:27220849) ![]() At higher concentrations (change in protein concentration) in vitro DAXX(495-740) forms condensed droplets however, this tendency is strongly enhanced in the presence of SPOP, and this was the case in the presence of both polymer and protein crowders, as evidenced by fluorescence microscopy of tagged protein constructs (PMID:30244836). In vitro studies demonstrated that SPOP(28-359) undergoes self-oligomerization (physical interaction), and in the presence of molecular crowders such as Ficoll-70, these oligomers are large enough to be observed by light microscopy (particle size and count). The co-expression of the two proteins change their localization, as SPOP localized to nuclear speckles, and DAXX localized to PML bodies when expressed alone. Different expression tags did not influence the co-localization of SPOP and DAXX. Transiently expressed mCherry-fused SPOP co-localized with GFP-fused DAXX (a SPOP substrate) in largely spherical type of nuclear bodies in vivo (morphology, protein localization) distinct from nuclear speckles, PML bodies, nucleoli, and Cajal bodies, as evidenced by flurescence microscopy. MSRVPSPPPPAEMSSGPVAESWCYTQIKVVKFSYMWTINNFSFCREEMGEVIKSSTFSSGANDKLKWCLRVNPKGLDEESKDYLSLYLLLVSCPKSEVRAKFKFSILNAKGEETKAMESQRAYRFVQGKDWGFKKFIRRDFLLDEANGLLPDDKLTLFCEVSVVQDSVNISGQNTMNMVKVPECRLADELGGLWENSRFTDCCLCVAGQEFQAHKAILAARSPVFSAMFEHEMEESKKNRVEINDVEPEVFKEMMCFIYTGKAPNLDKMADDLLAAADKYALERLKVMCEDALCSNLSVENAAEILILADLHSADQLKTQAVDFINYHASDVLETSGWKSMVVSHPHLVAEAYRSLASAQCPFLGPPRKRLKQS Rita Pancsa Bálint Mészáros Orsolya Kovács DAXX or androgen receptor (strictly required for LLPS) Morphological traits (PMID:30244836) rheological traits (PMID:30244836) dynamic movement/reorganization of molecules within the droplet assessed using FRAP (PMID:30244836)ġ) SPOP subtrates eg. 1) protein concentration of SPOP 2) substrate concentration 3) molar ratios SPOP:substrate 4) presence of cancer mutations (negatively affect LLPS)Ĭentral region containing the ordered MATH (substrate binding) and BTB domains (dimerization and CU元 binding)Īctivation/nucleation/signal amplification/bioreactor ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |