CO2-Responsive CCT Protein interacts with 14-3-3 proteins and controls
the expression of starch synthesis-related genes
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.