当前位置： SCI文献检索 > BMC BIOLOGY期刊下所有文献 > Two-component cyclase opsins of green algae are ATP-dependent and light-inhibited guanylyl cyclases.

Two-component cyclase opsins of green algae are ATP-dependent and light-inhibited guanylyl cyclases.

Abstract：

BACKGROUND:The green algae Chlamydomonas reinhardtii and Volvox carteri are important models for studying light perception and response, expressing many different photoreceptors. More than 10 opsins were reported in C. reinhardtii, yet only two-the channelrhodopsins-were functionally characterized. Characterization of new opsins would help to understand the green algae photobiology and to develop new tools for optogenetics. RESULTS:Here we report the characterization of a novel opsin family from these green algae: light-inhibited guanylyl cyclases regulated through a two-component-like phosphoryl transfer, called "two-component cyclase opsins" (2c-Cyclops). We prove the existence of such opsins in C. reinhardtii and V. carteri and show that they have cytosolic N- and C-termini, implying an eight-transmembrane helix structure. We also demonstrate that cGMP production is both light-inhibited and ATP-dependent. The cyclase activity of Cr2c-Cyclop1 is kept functional by the ongoing phosphorylation and phosphoryl transfer from the histidine kinase to the response regulator in the dark, proven by mutagenesis. Absorption of a photon inhibits the cyclase activity, most likely by inhibiting the phosphoryl transfer. Overexpression of Vc2c-Cyclop1 protein in V. carteri leads to significantly increased cGMP levels, demonstrating guanylyl cyclase activity of Vc2c-Cyclop1 in vivo. Live cell imaging of YFP-tagged Vc2c-Cyclop1 in V. carteri revealed a development-dependent, layer-like structure at the immediate periphery of the nucleus and intense spots in the cell periphery. CONCLUSIONS:Cr2c-Cyclop1 and Vc2c-Cyclop1 are light-inhibited and ATP-dependent guanylyl cyclases with an unusual eight-transmembrane helix structure of the type I opsin domain which we propose to classify as type Ib, in contrast to the 7 TM type Ia opsins. Overexpression of Vc2c-Cyclop1 protein in V. carteri led to a significant increase of cGMP, demonstrating enzyme functionality in the organism of origin. Fluorescent live cell imaging revealed that Vc2c-Cyclop1 is located in the periphery of the nucleus and in confined areas at the cell periphery.

journal_name

BMC Biol

journal_title

BMC biology

authors

Tian Y,Gao S,von der Heyde EL,Hallmann A,Nagel G

doi

10.1186/s12915-018-0613-5

subject

Has Abstract

pub_date

2018-12-06 00:00:00

pages

144

issue

issn

1741-7007

pii

10.1186/s12915-018-0613-5

journal_volume

pub_type

杂志文章

在线工具

Two-component cyclase opsins of green algae are ATP-dependent and light-inhibited guanylyl cyclases.