title: Dynamin controls extracellular level of Awd/Nme1 metastasis suppressor protein.
abstract: Dynamin GTPase (Dyn) plays a critical role in membrane-remodelling events underlying endocytosis. Studies in Drosophila identified a functional interaction between the Dyn homologue, encoded by the shibire (shi) gene, and Abnormal wing discs (Awd), a nucleoside diphosphate kinase (NDPK) that is the homologue of group I Nme human genes. These Drosophila studies showed that awd mutations enhance mutant shi phenotype and thus indicated the existence of a highly specific interaction between these genes. Furthermore, in human cells, it has been shown that Nme proteins promote Dyn activity in different membrane compartments through spatially controlled supply of GTP. Interestingly, Awd and Nme proteins have been detected in the extracellular environment. While no role has been inferred to extracellular Awd, presence of Nme1 in cancer patient serum is an unfavourable prognostic marker. In the present work, we used Drosophila and human cell line models to investigate the shuttling Awd/Nme1 proteins between intracellular and extracellular spaces. By using classic and reverse genetic approaches, we show that downregulation of Shi/Dyn1 activity enhances extracellular Awd/Nme1 in both Drosophila and human colon cell lines. We extended our analyses to colon cancer cell lines and found that knocking down Dyn1, besides to raise Nme1 extracellular amount, downregulates expression of molecular components that play key roles in tumour invasion. Interestingly, in vivo analyses of Drosophila larval adipocytes show that the conditional block of Shi activity greatly reduces intracellular amount of Awd confirming that Shi plays a key role in controlling the balance between intracellular and extracellular Awd.
authors.affiliation: Dipartimento di Farmacia e biotecnologie, Alma Mater Studiorum Università di Bologna, Bologna, Italy.|Dipartimento di Medicina molecolare, Università di Padova, Padova, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Romani
authors.affiliation: Dipartimento di Scienze biologiche, geologiche e ambientali, Alma Mater Studiorum Università di Bologna, Bologna, Italy.
authors.forename: Alessio
authors.initials: A
authors.lastname: Papi
authors.affiliation: Dipartimento di Farmacia e biotecnologie, Alma Mater Studiorum Università di Bologna, Bologna, Italy.
authors.forename: Marilena
authors.initials: M
authors.lastname: Ignesti
authors.affiliation: Dipartimento di Farmacia e biotecnologie, Alma Mater Studiorum Università di Bologna, Bologna, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Soccolini
authors.affiliation: Department of Medicine, Boston University School of Medicine, Boston, MA, USA.|Graduate Institute of Systems Biology and Bioinformatics, National Central University, Jhongli, Taiwan.
authors.forename: Tien
authors.initials: T
authors.lastname: Hsu
authors.affiliation: Dipartimento di Farmacia e biotecnologie, Alma Mater Studiorum Università di Bologna, Bologna, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Gargiulo
authors.affiliation: Dipartimento di Scienze biologiche, geologiche e ambientali, Alma Mater Studiorum Università di Bologna, Bologna, Italy. enzo.spisni@unibo.it.
authors.forename: Enzo
authors.initials: E
authors.lastname: Spisni
authors.affiliation: Dipartimento di Farmacia e biotecnologie, Alma Mater Studiorum Università di Bologna, Bologna, Italy. valeria.cavaliere@unibo.it.
authors.forename: Valeria
authors.identifier: http://orcid.org/0000-0002-3264-499X
authors.initials: V
authors.lastname: Cavaliere
mesh_terms: D017667:Adipocytes / Q000201:enzymology; D000818:Animals; D030801:Animals, Genetically Modified; D003110:Colonic Neoplasms / Q000201:enzymology* / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics; D034284:Dynamin I / Q000235:genetics / Q000378:metabolism*; D034281:Dynamins / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D005838:Genotype; D019073:HT29 Cells; D006801:Humans; D007814:Larva / Q000201:enzymology; D009154:Mutation; D054778:NM23 Nucleoside Diphosphate Kinases / Q000235:genetics / Q000378:metabolism*; D009701:Nucleoside-Diphosphate Kinase / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D034622:RNA Interference; D014162:Transfection
chemical_list: D029721:Drosophila Proteins; D054778:NM23 Nucleoside Diphosphate Kinases; C517938:NME1 protein, human; D009701:Nucleoside-Diphosphate Kinase; C061642:awd protein, Drosophila; D034284:Dynamin I; D034281:Dynamins; C439628:shi protein, Drosophila
keywords: Awd/Nme; Endocytosis; Shibire/Dynamin; Tumour suppressor gene