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CO2-Responsive CCT Protein interacts with 14-3-3 proteins and controls the expression of starch synthesis-related genes
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  • Hiroshi Fukayama,
  • Naoki Shibatani,
  • Hirofumi Miyagawa,
  • Aiko Koudou,
  • Yasuo Yamauchi,
  • Daisuke Matsuoka,
  • Daisuke Sasayama,
  • Tomoko Hatanaka,
  • Tetsushi Azuma,
  • Ryutaro Morita
Hiroshi Fukayama
Kobe University
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Naoki Shibatani
Kobe University
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Hirofumi Miyagawa
Kobe University
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Aiko Koudou
Kobe University
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Yasuo Yamauchi
Kobe University
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Daisuke Matsuoka
Kobe University
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Daisuke Sasayama
Kobe University
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Tomoko Hatanaka
Kobe University
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Tetsushi Azuma
Kobe University
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Ryutaro Morita
Kobe University
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Abstract

CO2 responsive CCT protein (CRCT) is a positive regulator of starch synthesis related genes such as ADP-glucose pyrophosphorylase large subunit 1 and starch branching enzyme I particularly in the leaf sheath of rice (Oryza sativa L.). The promoter GUS analysis revealed that CRCT expressed exclusively in the vascular bundle, whereas starch synthesis related genes were expressed in different sites such as mesophyll cell and starch storage parenchyma cell. However, the chromatin immunoprecipitation (ChIP) using a FLAG-CRCT overexpression line and subsequent qPCR analyses showed that the 5’-flanking regions of these starch synthesis-related genes tended to be enriched by ChIP, suggesting that CRCT can bind to the promoter regions of these genes. The monomer of CRCT is 34.2 kDa, however CRCT was detected at 270 kDa via gel filtration chromatography, suggesting that CRCT forms a complex in vivo. Immunoprecipitation and subsequent MS analysis pulled down several 14-3-3-like proteins. A yeast two-hybrid analysis and bimolecular fluorescence complementation assays confirmed the interaction between CRCT and 14-3-3-like proteins. Although there is an inconsistency in the place of expression, this study provide important findings regarding the molecular function of CRCT to control the expression of key starch synthesis-related genes.

Peer review status:ACCEPTED

29 Mar 2021Submitted to Plant, Cell & Environment
29 Mar 2021Submission Checks Completed
29 Mar 2021Assigned to Editor
29 Mar 2021Reviewer(s) Assigned
14 Apr 2021Review(s) Completed, Editorial Evaluation Pending
17 Apr 2021Editorial Decision: Revise Minor
23 Apr 20211st Revision Received
26 Apr 2021Submission Checks Completed
26 Apr 2021Assigned to Editor
02 May 2021Review(s) Completed, Editorial Evaluation Pending
02 May 2021Editorial Decision: Accept