title: A cyanobacterial light activated adenylyl cyclase partially restores development of a Dictyostelium discoideum, adenylyl cyclase a null mutant.
abstract: A light-regulated adenylyl cyclase, mPAC, was previously identified from the cyanobacterium Microcoleus chthonoplastes PCC7420. MPAC consists of a flavin-based blue light-sensing LOV domain and a catalytic domain. In this work, we expressed mPAC in an adenylate cyclase A null mutant (aca-) of the eukaryote Dictyostelium discoideum and tested to what extent light activation of mPAC could restore the cAMP-dependent developmental programme of this organism. Amoebas of Dictyostelium, a well-established model organism, generate and respond to cAMP pulses, which cause them to aggregate and construct fruiting bodies. mPAC was expressed under control of a constitutive actin-15 promoter in D. discoideum and displayed low basal adenylyl cyclase activity in darkness that was about five-fold stimulated by blue light. mPAC expression in aca- cells marginally restored aggregation and fruiting body formation in darkness. However, more and larger fruiting bodies were formed when mPAC expressing cells were incubated in light. Extending former applications of light-regulated AC, these results demonstrate that mPAC can be used to manipulate multicellular development in eukaryotes in a light dependent manner.
authors.affiliation: College of Life Sciences, University of Dundee, Dundee, UK.
authors.forename: Zhi-Hui
authors.initials: ZH
authors.lastname: Chen
authors.affiliation: MPI Chemical Energy Conversion, Mülheim, Germany.
authors.forename: Sarah
authors.initials: S
authors.lastname: Raffelberg
authors.affiliation: Department of Physics and Earth Sciences, University of Parma, Parma, Italy.
authors.forename: Aba
authors.initials: A
authors.lastname: Losi
authors.affiliation: College of Life Sciences, University of Dundee, Dundee, UK. Electronic address: p.schaap@dundee.ac.uk.
authors.forename: Pauline
authors.initials: P
authors.lastname: Schaap
authors.affiliation: MPI Chemical Energy Conversion, Mülheim, Germany. Electronic address: Wolfgang.gaertner@cec.mpg.de.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Gärtner
mesh_terms: D000262:Adenylyl Cyclases / Q000096:biosynthesis* / Q000235:genetics; D002449:Cell Aggregation / Q000235:genetics; D000458:Cyanobacteria / Q000201:enzymology*; D000242:Cyclic AMP / Q000235:genetics; D004023:Dictyostelium / Q000235:genetics* / Q000254:growth & development; D015971:Gene Expression Regulation, Enzymologic; D008027:Light; D009154:Mutation; D017434:Protein Structure, Tertiary / Q000235:genetics; D015398:Signal Transduction
chemical_list: D000242:Cyclic AMP; D000262:Adenylyl Cyclases
keywords: Cyclic AMP; LOV domain; Light-regulated enzyme activity; Optogenetics; Photoreceptors