MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it The Legionella Kinase LegK2 Targets the ARP2/3 Complex To Inhibit Actin Nucleation on Phagosomes and Allow Bacterial Evasion of the Late Endocytic Pathway. The Legionella Kinase LegK2 Targets the ARP2/3 Complex To Inhibit Actin Nucleation on Phagosomes and Allow Bacterial Evasion of the Late Endocytic Pathway. mBio 2015 imex curation Michard C., Sperandio D., Baïlo N., Pizarro-Cerdá J., LeClaire L., Chadeau-Argaud E., Pombo-Grégoire I., Hervet E., Vianney A., Gilbert C., Faure M., Cossart P., Doublet P. weiguohu@fudan.edu.cn 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Michard C., Sperandio D., Baïlo N., Pizarro-Cerdá J., LeClaire L., Chadeau-Argaud E., Pombo-Grégoire I., Hervet E., Vianney A., Gilbert C., Faure M., Cossart P., Doublet P. weiguohu@fudan.edu.cn Accepted 2018-MAR-05 AT 17:56 GMT AT 17:56 GMT by LUANA mBio 2015 Only protein-protein interactions imex curation imex curation The Legionella Kinase LegK2 Targets the ARP2/3 Complex To Inhibit Actin Nucleation on Phagosomes and Allow Bacterial Evasion of the Late Endocytic Pathway. The Legionella Kinase LegK2 Targets the ARP2/3 Complex To Inhibit Actin Nucleation on Phagosomes and Allow Bacterial Evasion of the Late Endocytic Pathway. mBio 2015 imex curation Michard C., Sperandio D., Baïlo N., Pizarro-Cerdá J., LeClaire L., Chadeau-Argaud E., Pombo-Grégoire I., Hervet E., Vianney A., Gilbert C., Faure M., Cossart P., Doublet P. weiguohu@fudan.edu.cn 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Michard C., Sperandio D., Baïlo N., Pizarro-Cerdá J., LeClaire L., Chadeau-Argaud E., Pombo-Grégoire I., Hervet E., Vianney A., Gilbert C., Faure M., Cossart P., Doublet P. weiguohu@fudan.edu.cn Accepted 2018-MAR-05 AT 17:56 GMT AT 17:56 GMT by LUANA mBio 2015 Only protein-protein interactions imex curation imex curation The Legionella Kinase LegK2 Targets the ARP2/3 Complex To Inhibit Actin Nucleation on Phagosomes and Allow Bacterial Evasion of the Late Endocytic Pathway. The Legionella Kinase LegK2 Targets the ARP2/3 Complex To Inhibit Actin Nucleation on Phagosomes and Allow Bacterial Evasion of the Late Endocytic Pathway. mBio 2015 imex curation Michard C., Sperandio D., Baïlo N., Pizarro-Cerdá J., LeClaire L., Chadeau-Argaud E., Pombo-Grégoire I., Hervet E., Vianney A., Gilbert C., Faure M., Cossart P., Doublet P. weiguohu@fudan.edu.cn 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. pull down pull down anti tag western anti tag western blot anti tag western Michard C., Sperandio D., Baïlo N., Pizarro-Cerdá J., LeClaire L., Chadeau-Argaud E., Pombo-Grégoire I., Hervet E., Vianney A., Gilbert C., Faure M., Cossart P., Doublet P. weiguohu@fudan.edu.cn Accepted 2018-MAR-05 AT 17:56 GMT AT 17:56 GMT by LUANA mBio 2015 Only protein-protein interactions imex curation imex curation arp3_human Actin-related protein 3 ACTR3 ARP3 Actin-like protein 3 protein protein human Homo sapiens Human MAGRLPACVVDCGTGYTKLGYAGNTEPQFIIPSCIAIKESAKVGDQAQRRVMKGVDDLDFFIGDEAIEKPTYATKWPIRHGIVEDWDLMERFMEQVIFKYLRAEPEDHYFLLTEPPLNTPENREYTAEIMFESFNVPGLYIAVQAVLALAASWTSRQVGERTLTGTVIDSGDGVTHVIPVAEGYVIGSCIKHIPIAGRDITYFIQQLLRDREVGIPPEQSLETAKAVKERYSYVCPDLVKEFNKYDTDGSKWIKQYTGINAISKKEFSIDVGYERFLGPEIFFHPEFANPDFTQPISEVVDEVIQNCPIDVRRPLYKNIVLSGGSTMFRDFGRRLQRDLKRTVDARLKLSEELSGGRLKPKPIDVQVITHHMQRYAVWFGGSMLASTPEFYQVCHTKKDYEEIGPSICRHNPVFGVMS DIP protein P61158 original sequence version: 117 DIP protein P61158 original sequence version: 84 DIP protein P61158 original sequence version: 127 23E5564198B81C63 q5ztm4_legph legK2 lpg2137 protein protein legph Legionella pneumophila subsp. pneumophila MVYYINLKEQPLHQDKEILTQLIKQYLLSGSSQSALFANHPYTLNDEDGNEIEFLLSHDIYKNQREDGITEFLILNGSQIGRGTKGVVIQSNHAMYLIDDELVIFPKENVLKVLYQNFINAEELIKQESVNYSILSGNSTAELVNLQGGNQGFFMQKEEGYALEELLEDIDSLQKLTFSDKMELAISFLTELENLHNKGYAHGDLTFKNILYNEKNNQLKIIDAGEMKVADERFWAQDISIAIEMISLLFTTNETNQFKASRYWSDEPTVRRIVNPNPVEAKLNRILNSEYRCVSDLKLAIERNYNEYLNNSNLVLKKNVELESEALGLQAYKNQMSKQQLLGEIQGFKENYWNMKDLLTLTNRHHLRVFLEYLDNICSAFKDDKTDEKSARAAYDFLNAQINKLFEDNSKNSKPSFESFSEDVQRFLIHIDTYLMKNPSACSNSIASTIQLLKQLDNKKSFNPEQSFKDFCSYKEITIQLLLKPFETPVAEMASAPAIASDLIEKQSDTKRKLQEMKSSANQDKQPIFTPSLMRGPS F086A8AD1E68515B arc1b_human Actin-related protein 2/3 complex subunit 1B Arp2/3 complex 41 kDa subunit p41-ARC ARPC1B ARC41 protein protein human Homo sapiens Human MAYHSFLVEPISCHAWNKDRTQIAICPNNHEVHIYEKSGAKWTKVHELKEHNGQVTGIDWAPESNRIVTCGTDRNAYVWTLKGRTWKPTLVILRINRAARCVRWAPNENKFAVGSGSRVISICYFEQENDWWVCKHIKKPIRSTVLSLDWHPNNVLLAAGSCDFKCRIFSAYIKEVEERPAPTPWGSKMPFGELMFESSSSCGWVHGVCFSASGSRVAWVSHDSTVCLADADKKMAVATLASETLPLLALTFITDNSLVAAGHDCFPVLFTYDAAAGMLSFGGRLDVPKQSSQRGLTARERFQNLDKKASSEGGTAAGAGLDSLHKNSVSQISVLSGGKAKCSQFCTTGMDGGMSIWDVKSLESALKDLKIK DIP protein O15143 original sequence version: 110 DIP protein O15143 original sequence version: 150 1939F5B63D40BD27 actr3-arpc1b-1 2 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 5 predetermined predetermined participant predetermined nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell physical association physical association f1a Reproducibly, several candidates for interaction with LegK2 were proteins involved in actin cytoskeleton assembly. Because the legK2 mutant phenotype alters host cell vesicle trafficking during infection, particular attention was given to proteins of this family, which included ARP3 and ARPC1B subunits of the actin filament nucleator ARP2/3 complex. arpc1b-legk2-1 3 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell gst tag gst tag 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 physical association physical association f1b As ARPC1B was not purified with GST, these data confirm that LegK2 specifically interacts with the ARPC1B subunit of the ARP2/3 complex in mammalian cells. Regarding ARP3, we propose that the interaction between LegK2 and ARP3 is too weak and/or transient to be detected by affinity copurification under our conditions actr3-arpc1b-2 1 4 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell flag tag flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell gst tag gst tag 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 colocalization colocalization f1c Both antibodies labeled the cytoplasm and periphery of cells with similar staining patterns when cells were cotransfected with pDEST27-legK2 and pCI-Neo3Flag-ACTR3 or -ARPC1B, while anti-GST labeling in cells cotransfected with the empty vector or the LegK1 expressing vector was diffused in the cell. These observations suggest that LegK2 specifically colocalizes with ARPC1B and ARP3. legk2-actr3-1 2 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 5 predetermined predetermined participant predetermined nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell physical association physical association f1a Reproducibly, several candidates for interaction with LegK2 were proteins involved in actin cytoskeleton assembly. Because the legK2 mutant phenotype alters host cell vesicle trafficking during infection, particular attention was given to proteins of this family, which included ARP3 and ARPC1B subunits of the actin filament nucleator ARP2/3 complex. arpc1b-legk2-2 1 6 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell flag tag flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell gst tag gst tag 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 colocalization colocalization f1c Both antibodies labeled the cytoplasm and periphery of cells with similar staining patterns when cells were cotransfected with pDEST27-legK2 and pCI-Neo3Flag-ACTR3 or -ARPC1B, while anti-GST labeling in cells cotransfected with the empty vector or the LegK1 expressing vector was diffused in the cell. These observations suggest that LegK2 specifically colocalizes with ARPC1B and ARP3.