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Biochim Biophys Acta. 2013 Dec;1833(12):2775-2780. doi: 10.1016/j.bbamcr.2013.06.024. Epub 2013 Jul 16.

A low temperature-inducible protein AtSRC2 enhances the ROS-producing activity of NADPH oxidase AtRbohF.

Author information

1
Department of Applied Biological Science, Tokyo University of Science, Noda, Chiba 278-8510, Japan.
2
Department of Biological Science, Graduate School of Science, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
3
Department of Applied Biological Science, Tokyo University of Science, Noda, Chiba 278-8510, Japan; Department of Biological Science, Graduate School of Science, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan. Electronic address: hidetaka@biol.s.u-tokyo.ac.jp.
4
Department of Applied Biological Science, Tokyo University of Science, Noda, Chiba 278-8510, Japan. Electronic address: kuchitsu@rs.noda.tus.ac.jp.

Abstract

Reactive oxygen species (ROS) produced by NADPH oxidases play critical roles in plant environmental responses. Arabidopsis thaliana NADPH oxidase AtRbohF-mediated ROS-production is involved in abiotic stress responses. Because overproduction of ROS is highly toxic to cells, the activity of AtRbohF needs to be tightly regulated in response to diverse stimuli. The ROS-producing activity of AtRbohF is activated by Ca(2+) and protein phosphorylation, but other regulatory factors for AtRbohF are mostly unknown. In this study, we screened for proteins that interact with the N-terminal cytosolic region of AtRbohF by a yeast two-hybrid screen, and isolated AtSRC2, an A. thaliana homolog of SRC2 (soybean gene regulated by cold-2). A co-immunoprecipitation assay revealed that AtSRC2 interacts with the N-terminal region of AtRbohF in plant cells. Intracellular localization of GFP-tagged AtSRC2 was partially overlapped with that of GFP-tagged AtRbohF at the cell periphery. Co-expression of AtSRC2 enhanced the Ca(2+)-dependent ROS-producing activity of AtRbohF in HEK293T cells, but did not affect its phosphorylation-dependent activation. Low-temperature treatment induced expression of the AtSRC2 gene in Arabidopsis roots in proportion to levels of ROS production that was partially dependent on AtRbohF. Our findings suggest that AtSRC2 is a novel activator of Ca(2+)-dependent AtRbohF-mediated ROS production and may play a role in cold responses.

KEYWORDS:

ABA; Arabidopsis thaliana; Arabidopsis thaliana respiratory burst oxidase homologue; AtRboh; AtRbohF; Cold stress; NADPH oxidase; NADPH oxidase (NOX); NOX; ROS; RT-PCR; Reactive oxygen species (ROS); Respiratory burst oxidase homologue (Rboh); SRC2; TM; abscisic acid; co-IP; co-immunoprecipitation; reactive oxygen species; reverse transcription-polymerase chain reaction; soybean gene regulated by cold-2; transmembrane

PMID:
23872431
DOI:
10.1016/j.bbamcr.2013.06.024
[Indexed for MEDLINE]
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