UniProt UniProt http://www.uniprot.org Cortical localization of the Galpha protein GPA-16 requires RIC-8 function during C. elegans asymmetric cell division. Cortical localization of the Galpha protein GPA-16 requires RIC-8 function during C. elegans asymmetric cell division. Development (0950-1991) 2005 Afshar K., Willard FS., Colombo K., Siderovski DP., Gonczy P. The proteins gpr-1 and/or gpr-2 may also interact with goa-1 and gpa-16; due to the high degree of sequence similarity, these two proteins cannot be distinguished in coimmunoprecipitation experiments. pierre.gonczy@isrec.unil.ch in vitro In vitro biochemical biochemical predetermined predetermined participant predetermined Afshar K., Willard FS., Colombo K., Siderovski DP., Gonczy P. The proteins gpr-1 and/or gpr-2 may also interact with goa-1 and gpa-16; due to the high degree of sequence similarity, these two proteins cannot be distinguished in coimmunoprecipitation experiments. Time-course of [35S]GTP{gamma}S binding to 100 nM GPA-16 in the presence or absence of 10 �M GPR-1/2 peptide studied. pierre.gonczy@isrec.unil.ch Accepted 2006-OCT-9 by JYOTI. Development (0950-1991) 2005 Cortical localization of the Galpha protein GPA-16 requires RIC-8 function during C. elegans asymmetric cell division. Cortical localization of the Galpha protein GPA-16 requires RIC-8 function during C. elegans asymmetric cell division. Development (0950-1991) 2005 Afshar K., Willard FS., Colombo K., Siderovski DP., Gonczy P. The proteins gpr-1 and/or gpr-2 may also interact with goa-1 and gpa-16; due to the high degree of sequence similarity, these two proteins cannot be distinguished in coimmunoprecipitation experiments. pierre.gonczy@isrec.unil.ch in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Afshar K., Willard FS., Colombo K., Siderovski DP., Gonczy P. The proteins gpr-1 and/or gpr-2 may also interact with goa-1 and gpa-16; due to the high degree of sequence similarity, these two proteins cannot be distinguished in coimmunoprecipitation experiments. pierre.gonczy@isrec.unil.ch Accepted 2006-OCT-9 by JYOTI. Development (0950-1991) 2005 Cortical localization of the Galpha protein GPA-16 requires RIC-8 function during C. elegans asymmetric cell division. Cortical localization of the Galpha protein GPA-16 requires RIC-8 function during C. elegans asymmetric cell division. Development (0950-1991) 2005 Afshar K., Willard FS., Colombo K., Siderovski DP., Gonczy P. The proteins gpr-1 and/or gpr-2 may also interact with goa-1 and gpa-16; due to the high degree of sequence similarity, these two proteins cannot be distinguished in coimmunoprecipitation experiments. pierre.gonczy@isrec.unil.ch caeel-embryo Caenorhabditis elegans embryo embryo Embryo Defined as the development stage prior to the initiation of tissue differentiation. anti bait coip anti bait coimmunoprecipitation anti bait coip polyclonal western polyclonal antibody western blot polyclonal western Afshar K., Willard FS., Colombo K., Siderovski DP., Gonczy P. The proteins gpr-1 and/or gpr-2 may also interact with goa-1 and gpa-16; due to the high degree of sequence similarity, these two proteins cannot be distinguished in coimmunoprecipitation experiments. pierre.gonczy@isrec.unil.ch Accepted 2006-OCT-9 by JYOTI. Development (0950-1991) 2005 gnao_caeel Guanine nucleotide-binding protein G(o) subunit alpha goa-1 C26C6.2 protein protein caeel Caenorhabditis elegans MGCTMSQEERAALERSRMIEKNLKEDGMQAAKDIKLLLLGAGESGKSTIVKQMKIIHESGFTAEDYKQYKPVVYSNTVQSLVAILRAMSNLGVSFGSADREVDAKLVMDVVARMEDTEPFSEELLSSMKRLWGDAGVQDCFSRSNEYQLNDSAKYFLDDLERLGEAIYQPTEQDILRTRVKTTGIVEVHFTFKNLNFKLFDVGGQRSERKKWIHCFEDVTAIIFCVAMSEYDQVLHEDETTNRMHESLKLFDSICNNKWFTDTSIILFLNKKDLFEEKIKKSPLTICFPEYSGRQDYHEASAYIQAQFEAKNKSANKEIYCHMTCATDTTNIQFVFDAVTDVIIANNLRGCGLY 8C98245C8E9C48B0 gpa16_caeel Guanine nucleotide-binding protein alpha-16 subunit gpa-16 spn-1 Y95B8A.5 protein protein caeel Caenorhabditis elegans MGCIMSQEDEAAKRRSKKIDRLLKEDGENSMRTIKLLLLGAGESGKSTILKQMRIIHDVGYTTEERKVFRGVVYGNIILSLNAIIHAMEQLKISFTTLDHESDARKLLMFSTTGEEDELPEELVVLMKSVWSDSGIQKALERSREYQLNDSAGYYLSQLDRICAPNYIPTQDDILRTRIKTTGIVETQFVYKDRLFLVFDVGGQRSERKKWIHCFEDVTALIFCVALSEYDMVLVEDCQTNRMRESLKLFDSICNNKWFVETSIILFLNKKDLFEEKIVRSPLTHCFPEYTGANNYEEASAYIQQQFEDMNKRTTGEKNQEIYTQFTCATDTNNIRFVFDAVTDIIIRDNLRTCGLY C0CDB733D06EE6C9 q9gsx9-1 Synembryn ric-8 Resistance to inhibitors of cholinesterase protein 8 Y69A2AR.2 protein protein caeel Caenorhabditis elegans MSEELHSDLIASIFGGKPAKIEEFFSKWNFANAAVSKFDMANSAKNELGDRICEVIENGELTHVLLETIKILSREKDGLEGLLNDPLCDKILAFAELSSNENNSKTVHTLMEAQKCLINTLFHSQRMRDRFYANPKTGENLQFFLGEFEENRRKTSSIDWIRLLNPVQAAEIWYFYHRIAFIATALGREFQKNWANDPKTIDSLLLAVEICTNRSENSTQDINRATEALKTFFNVFCHFHGDVKAIDHKNAAKTCQILRDAICSDVLTDDVVQSAIHCLSVPPLPMVLSVLCGKNSKNNGGENEEEKFFVEELSNMQLTEAILMHLDKQLTKVVALLLNDAPNQQQNPMLSAEASTLTDLVGPYFQVLARLCTDSKYVRRYCRIRVIPPLVSEEVQKRPEENNTLRGRIARIMMLPSSTKDVAAEFLFIICKRSVNRMIKYLGFGHSAGHLANLGLLGQINQPKHASDSEDSETEDYNQIKDSVNPVTGAIYPSDHGSALAGMSEEQKEYEAMKLVDAMNQMMETGIVKPGTIGDDGKIREVSHVLELLKNAPEPAPAENSDSDEE 10C4E5423BB81FBF gpr1_caeel G-protein regulator 1 F22B7.13 gpr-1 protein protein caeel Caenorhabditis elegans MDVSYYDGPKDEVAEAMLKSAVTAMRLGQYEDGKGRLEEIMEFGTSNFQLLGTIYMYYGRVCRHLNHDAKALEFFEHELNMFKLIFNYPEACDSTRRIVEQALKMGKFPKARRFAEDLIDYTSNKKNGEKYIGQARILFASVCLEGCERDVESNQDEKKKLLSICAEQIAAVKLFNENNTEGAVSETKIMLLEAKCLSLDEKYEESRRKYQECIDFAIKTDQFEAVHIAYYDKALYAETDLLFFIIRDLRSALFYATKFGKERDVVKYKSKLSEEMLRNGEFHEAYLYGLEALVSIRKLGLNEYIGDVLLTIAKCLIALGKRRQAAYFIILGSVLTINQNSFKLFYEQIDVAMNQERSETATDQDVCLAIDSSPDPTSSNDMINKFVVELEHATNVETWEMIVNGIIDDQKKPVAIEKKENEEPVDMMDLIFSMSSRMDDQRTELPAARFIPPRPVSSASKKTTKSHRILPGLRANWTKVQSMKFDGHTMNRILKRSKKSKSSLDSTNSMQGDDTRSDDVTMTSK EC68F930E3622AB5 gpa-16_ric-8_1 3 5 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association Fig. 2A, 5G gpr-1_gpa-16_1 2 5 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range trini Trichoplusia ni (Cabbage looper) Cabbage looper 7 unspecified role unspecified role bait bait n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association This interaction is stimulated by GDP binding to gpa-16. Fig. 5A ric-8_gpa-16_1 2 6 unspecified role unspecified role bait bait n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain trini Trichoplusia ni (Cabbage looper) Cabbage looper physical association physical association This interaction is stimulated by GDP binding to gpa-16. Fig. 5B ric-8_goa-1_6 2 6 unspecified role unspecified role bait bait n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association This interaction is stimulated by GDP binding to goa-1. Fig. 5E goa-1_ric-8_2 3 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association This interaction is inhibited by GTP-gamma S. Fig. 5H gpr-1_gpa-16_2 1 5 unspecified role unspecified role neutral component neutral component n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain trini Trichoplusia ni (Cabbage looper) Cabbage looper 7 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 5C goa-1_ric-8_3 1 6 unspecified role unspecified role neutral component neutral component n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role neutral component neutral component n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association Fig. 5F