A light-regulated adenylyl cyclase, mPAC, was previously identified from the cyanobacterium Microcoleuschthonoplastes PCC7420. MPAC consists of a flavin-based blue light-sensing LOV domain and a catalyticdomain. In this work, we expressed mPAC in an adenylate cyclase A null mutant (aca−) of the eukaryoteDictyostelium 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-establishedmodel organism, generate and respond to cAMP pulses, which cause them to aggregate and constructfruiting bodies. mPAC was expressed under control of a constitutive actin-15 promoter in D. discoideumand displayed low basal adenylyl cyclase activity in darkness that was about five-fold stimulated byblue light. mPAC expression in aca− cells marginally restored aggregation and fruiting body formationin darkness. However, more and larger fruiting bodies were formed when mPAC expressing cells wereincubated in light. Extending former applications of light-regulated AC, these results demonstrate thatmPAC can be used to manipulate multicellular development in eukaryotes in a light dependent manner.

A cyanobacterial light activated adenylyl cyclase partially restores development of a Dictyostelium discoideum, adenylyl cyclase a null mutant / Zhi-hui Chen;Sarah Raffelberg;Aba Losi;Pauline Schaap;Wolfgang Gärtner. - In: JOURNAL OF BIOTECHNOLOGY. - ISSN 0168-1656. - 191(2014), pp. 246-249. [10.1016/j.jbiotec.2014.08.008]

A cyanobacterial light activated adenylyl cyclase partially restores development of a Dictyostelium discoideum, adenylyl cyclase a null mutant

LOSI, Aba;
2014

Abstract

A light-regulated adenylyl cyclase, mPAC, was previously identified from the cyanobacterium Microcoleuschthonoplastes PCC7420. MPAC consists of a flavin-based blue light-sensing LOV domain and a catalyticdomain. In this work, we expressed mPAC in an adenylate cyclase A null mutant (aca−) of the eukaryoteDictyostelium 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-establishedmodel organism, generate and respond to cAMP pulses, which cause them to aggregate and constructfruiting bodies. mPAC was expressed under control of a constitutive actin-15 promoter in D. discoideumand displayed low basal adenylyl cyclase activity in darkness that was about five-fold stimulated byblue light. mPAC expression in aca− cells marginally restored aggregation and fruiting body formationin darkness. However, more and larger fruiting bodies were formed when mPAC expressing cells wereincubated in light. Extending former applications of light-regulated AC, these results demonstrate thatmPAC can be used to manipulate multicellular development in eukaryotes in a light dependent manner.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11381/2781929
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