bims-plasge Biomed News
on Plastid genes
Issue of 2020‒05‒31
one paper selected by
Vera S. Bogdanova
Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences

  1. J Biol Chem. 2020 May 27. pii: jbc.RA120.012877. [Epub ahead of print]
      Acetyl-CoA carboxylase (ACCase) catalyzes the first committed step in de novo synthesis of fatty acids. The multisubunit ACCase in the chloroplast is activated by a shift to pH 8 upon light adaptation and is inhibited by a shift to pH 7 upon dark adaptation. Here, titrations with the purified ACCase BADC and BCCP subunits from Arabidopsis indicated that they can competently and independently bind biotin carboxylase (BC), but differ in responses to pH changes representing those in the plastid stroma during light or dark conditions. At pH 7 in phosphate buffer, BADC1 and BADC2 gain an advantage over BCCP1 and BCCP2 in affinity for BC. At pH 8 in KCl solution, however, BCCP1 and BCCP2 had more than 10-fold higher affinity for BC than did BADC1. The pH-modulated shifts in BC preferences for BCCP and BADC partners suggest they contribute to light-dependent regulation of heteromeric ACCase. Using NMR spectroscopy, we found evidence for increased intrinsic disorder of the BADC and BCCPs subunits at pH 7. We propose that this intrinsic disorder potentially promotes fast association with BC through a "fly-casting mechanism." We hypothesize that the pH effects on the BADC and BCCP subunits attenuate ACCase activity by night and enhance it by day. Consistent with this hypothesis, Arabidopsis badc1 badc3 mutant lines grown in a light-dark cycle synthesized more fatty acids in their seeds. In summary, our findings provide evidence that the BADC and BCCP subunits function as pH sensors required for light-dependent switching of heteromeric ACCase activity.
    Keywords:  Arabidopsis; biophysics; fatty acid biosynthesis; intrinsically disordered protein; lipid synthesis; nuclear magnetic resonance (NMR); pH regulation; plant biochemistry; protein-protein interaction; thermodynamics