New Drosophila circadian clock mutants affecting temperature compensation induced by targeted mutagenesis of timeless

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60077344%3A_____%2F19%3A00517191" target="_blank" >RIV/60077344:_____/19:00517191 - isvavai.cz</a>

Alternative codes found

RIV/60076658:12310/19:43899762

Result on the web

<a href="https://www.frontiersin.org/articles/10.3389/fphys.2019.01442/full" target="_blank" >https://www.frontiersin.org/articles/10.3389/fphys.2019.01442/full</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3389/fphys.2019.01442" target="_blank" >10.3389/fphys.2019.01442</a>

Result language

angličtina

Original language name

New Drosophila circadian clock mutants affecting temperature compensation induced by targeted mutagenesis of timeless

Original language description

Drosophila melanogaster has served as an excellent genetic model to decipher the molecular basis of the circadian clock. Two key proteins, PERIOD (PER) and TIMELESS (TIM), are particularly well explored and a number of various arrhythmic, slow, and fast clock mutants have been identified in classical genetic screens. Interestingly, the free running period (tau, τ) is influenced by temperature in some of these mutants, whereas τau is temperature-independent in other mutant lines as in wild-type flies. This, so-called “temperature compensation” ability is compromised in the mutant timeless allele “ritsu” (tim(rit), and, as we show here, also in the tim(blind) allele, mapping to the same region of TIM. To test if this region of TIM is indeed important for temperature compensation, we generated a collection of new mutants and mapped functional protein domains involved in the regulation of τau and in general clock function. We developed a protocol for targeted mutagenesis of specific gene regions utilizing the CRISPR/Cas9 technology, followed by behavioral screening. In this pilot study, we identified 20 new timeless mutant alleles with various impairments of temperature compensation. The mutations included short in-frame insertions, deletions, or substitutions of a few amino acids resulting from the non-homologous end joining repair process. Interestingly, several mutations with a strong temperature compensation defect map to one specific region of TIM. Although the exact mechanism of how these mutations affect TIM function is as yet unknown, our in silico analysis suggests they affect a putative nuclear export signal (NES) and phosphorylation sites of TIM. Immunostaining for PER was performed on two TIM mutants that display longer τau at 25°C and complete arrhythmicity at 28°C. Consistently with the behavioral phenotype, PER immunoreactivity was reduced in circadian clock neurons of flies exposed to elevated temperatures.

Czech name

—

Czech description

—

Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10602 - Biology (theoretical, mathematical, thermal, cryobiology, biological rhythm), Evolutionary biology

Project

<a href="/en/project/GA17-01003S" target="_blank" >GA17-01003S: Insect Photoperiodic Timer</a><br>

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2019

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Frontiers in physiology

ISSN

1664-042X

e-ISSN

—

Volume of the periodical

10

Issue of the periodical within the volume

DEC 03

Country of publishing house

CH - SWITZERLAND

Number of pages

23

Pages from-to

1442

UT code for WoS article

000503031900001

EID of the result in the Scopus database

2-s2.0-85077245619