A novel activation domain is essential for CIA5-mediated gene regulation in response to CO2 changes in Chlamydomonas reinhardtii

Chen, Bo; Lee, Kwanghong; Plucinak, Tom; Duanmu, Deqiang; Wang, Yingjun; Horken, Kempton M.; Weeks, Donald P.; Spalding, Martin H.

doi:10.1016/j.algal.2017.03.006

A novel activation domain is essential for CIA5-mediated gene regulation in response to CO₂ changes in Chlamydomonas reinhardtii

Journal Article · Tue Apr 04 00:00:00 EDT 2017 · Algal Research

DOI:https://doi.org/10.1016/j.algal.2017.03.006· OSTI ID:1533501

^[1]; Lee, Kwanghong ^[2]; Plucinak, Tom ^[3]; ^[4]; Wang, Yingjun ^[5]; Horken, Kempton M. ^[6]; Weeks, Donald P. ^[6]; ^[5]

Iowa State University, Ames, IA (United States); DOE/OSTI
University of Nebraska, Lincoln, NE (United States); Korea Seed & Variety Service, Miryang (Korea, Republic of)
University of Nebraska, Lincoln, NE (United States); GlaxoSmithKline, Research Triangle Park, NC (United States)
Iowa State University, Ames, IA (United States); Huazhong Agricultural University, Wuhan (China)
Iowa State University, Ames, IA (United States)
University of Nebraska, Lincoln, NE (United States)

The inducible CO₂ concentrating mechanism (CCM) of microalgae is essential for their acclimation to highly variable aquatic inorganic carbon levels. The key transcriptional regulator, CIA5, affects expression of thousands of genes in Chlamydomonas reinhardtii, but the molecular characteristics of this important protein are poorly understood. Here this study identifies a functional activation domain in CIA5 and demonstrates the functionality of a mini-CIA5 including only the zinc-binding domain and the identified activation domain. A highly conserved 130aa region from CIA5 exhibits auto-activation in yeast and appears responsible for the markedly slow migration of CIA5 when analyzed by SDS-PAGE. This 130aa region or either half of this region also effectively replaced the activation domain of a modified designer Transcription Activator-like Element (dTALE) in targeted activation of an endogenous Chlamydomonas gene. Additionally, a mini-CIA5 combining the conserved zinc-binding domain with the 130aa putative activation domain complemented the growth phenotype of the cia5 mutant and triggered CO₂-regulated gene expression patterns similar to wild-type cells or cia5 complemented with the full-length CIA5. Although the mini-CIA5 complementation did not fully restore wild-type growth rates or full gene induction/repression amplitudes, especially in very low CO₂, this newly identified activation domain combined with the previously described zinc-binding domain are demonstrated to be the key essential components of CIA5 that permit rapid CIA5-mediated responses to changes in CO₂ concentrations.

View Accepted Manuscript (DOE)

Research Organization:: University of Nebraska, Lincoln, NE (United States); University of California, San Diego, CA (United States)

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE); National Science Foundation (NSF); National Natural Science Foundation of China

Grant/Contract Number:: EE0001052; EE0003373

OSTI ID:: 1533501

Journal Information:: Algal Research, Journal Name: Algal Research Journal Issue: PA Vol. 24; ISSN 2211-9264

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CCM
CIA5
CO2 concentrating mechanism
Chlamydomonas
activation domain
recombinant mini-CIA5