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CLOCKS DO NOT TICK IN UNISON: ISOLATION OF CLOCK AND VRILLE SHED NEW LIGHT ON THE CLOCKWORK MODEL OF THE SAND FLY LUTZOMYIA LONGIPALPIS
Circadian clocks
Clock, vrille
PAS domain
bZIP domain
bHLH domain
Visceral leishmaniasis
Author
Affilliation
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Mosquitos Vetores: Endossimbiontes e Interação Patógeno Vetor. Belo Horizonte, MG, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Epidemiologia e Sistemática Molecular. Rio de Janeiro, RJ, Brasil.
École Polytechnique Fédérale de Lausanne. School of Life Sciences. , Institute of Bioengineering. Laboratory of Systems Biology and Genetics. Lausanne, Switzerland.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório Transmissores de Leishmanioses. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM)/ CNPq. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM)/ CNPq. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Epidemiologia e Sistemática Molecular. Rio de Janeiro, RJ, Brasil.
École Polytechnique Fédérale de Lausanne. School of Life Sciences. , Institute of Bioengineering. Laboratory of Systems Biology and Genetics. Lausanne, Switzerland.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório Transmissores de Leishmanioses. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM)/ CNPq. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM)/ CNPq. Rio de Janeiro, RJ, Brasil.
Abstract
Background: Behavior rhythms of insect vectors directly interfere with the dynamics of pathogen transmission to
humans. The sand fly Lutzomyia longipalpis is the main vector of visceral leishmaniasis in America and concentrates
its activity around dusk. Despite the accumulation of behavioral data, very little is known about the molecular bases
of the clock mechanism in this species. This study aims to characterize, within an evolutionary perspective, two
important circadian clock genes, Clock and vrille.
Findings: We have cloned and isolated the coding sequence of L. longipalpis’ genes Clock and vrille. The former is
structured in eight exons and encodes a protein of 696 amino acids, and the latter comprises three exons and translates
to a protein of 469 amino acids. When compared to other insects’ orthologues, L. longipalpis CLOCK shows a high
degree of conservation in the functional domains bHLH and PAS, but a much shorter glutamine-rich (poly-Q) C-terminal
region. As for L. longipalpis VRILLE, a high degree of conservation was found in the bZIP domain. To support these
observations and provide an elegant view of the evolution of both genes in insects, phylogenetic analyses based on
maximum-likelihood and Bayesian inferences were performed, corroborating the previously known insect systematics.
Conclusions: The isolation and phylogenetic analyses of Clock and vrille orthologues in L. longipalpis bring novel and
important data to characterize this species’ circadian clock. Interestingly, the poly-Q shortening observed in CLOCK
suggests that its transcription activity might be impaired and we speculate if this effect could be compensated by other
clock factors such as CYCLE.
Keywords
Lutzomyia longipalpiCircadian clocks
Clock, vrille
PAS domain
bZIP domain
bHLH domain
Visceral leishmaniasis
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