Methylcrotonyl-CoA Carboxylase Regulates Triacylglycerol Accumulation in the Model Diatom Phaeodactylum tricornutum.
The model marine diatom Phaeodactylum tricornutum can accumulate high levels of triacylglycerols (TAGs) under nitrogen depletion and has attracted increasing attention as a potential system for biofuel production. However, the molecular mechanisms involved in TAG accumulation in diatoms are largely unknown. Here, we employed a label-free quantitative proteomics approach to estimate differences in protein abundance before and after TAG accumulation. We identified a total of 1193 proteins, 258 of which were significantly altered during TAG accumulation. Data analysis revealed major changes in proteins involved in branched-chain amino acid (BCAA) catabolic processes, glycolysis, and lipid metabolic processes. Subsequent quantitative RT-PCR and protein gel blot analysis confirmed that four genes associated with BCAA degradation were significantly upregulated at both the mRNA and protein levels during TAG accumulation. The most significantly upregulated gene, encoding the β-subunit of methylcrotonyl-CoA carboxylase (MCC2), was selected for further functional studies. Inhibition of MCC2 expression by RNA interference disturbed the flux of carbon (mainly in the form of leucine) toward BCAA degradation, resulting in decreased TAG accumulation. MCC2 inhibition also gave rise to incomplete utilization of nitrogen, thus lowering biomass during the stationary growth phase. These findings help elucidate the molecular and metabolic mechanisms leading to increased lipid production in diatoms.
Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Feng
F
Ge
Diatom Biology Group, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Weichao
W
Huang
Diatom Biology Group, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Zhuo
Z
Chen
Diatom Biology Group, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Chunye
C
Zhang
Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Qian
Q
Xiong
Environmental and Evolutionary Genomics Section, Institut de Biologie de l'Ecole Normale Supérieure, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 8197, Institut National de la Santé et de la Recherche Médicale U1024, Ecole Normale Supérieure, 75230 Paris cedex 05, France.
Chris
C
Bowler
Diatom Biology Group, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Juan
J
Yang
Diatom Biology Group, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Jin
J
Xu
Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China Diatom Biology Group, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China hanhuahu@ihb.ac.cn.
Hanhua
H
Hu