Posttranslational regulation of pyruvate, orthophosphate dikinase in developing rice (Oryza sativa) seeds.
Pyruvate, orthophosphate dikinase (PPDK; E.C.2.7.9.1) is most well known as a photosynthetic enzyme in C4 plants. The enzyme is also ubiquitous in C3 plant tissues, although a precise non-photosynthetic C3 function(s) is yet to be validated, owing largely to its low abundance in most C3 organs. The single C3 organ type where PPDK is in high abundance, and, therefore, where its function is most amenable to elucidation, are the developing seeds of graminaceous cereals. In this report, we suggest a non-photosynthetic function for C3 PPDK by characterizing its abundance and posttranslational regulation in developing Oryza sativa (rice) seeds. Using primarily an immunoblot-based approach, we show that PPDK is a massively expressed protein during the early syncitial-endosperm/-cellularization stage of seed development. As seed development progresses from this early stage, the enzyme undergoes a rapid, posttranslational down-regulation in activity and amount via regulatory threonyl-phosphorylation (PPDK inactivation) and protein degradation. Immunoblot analysis of separated seed tissue fractions (pericarp, embryo + aleurone, seed embryo) revealed that regulatory phosphorylation of PPDK occurs in the non-green seed embryo and green outer pericarp layer, but not in the endosperm + aleurone layer. The modestly abundant pool of inactive PPDK (phosphorylated + dephosphorylated) that was found to persist in mature rice seeds was shown to remain largely unchanged (inactive) upon seed germination, suggesting that PPDK in rice seeds function in developmental rather than in post-developmental processes. These and related observations lead us to postulate a putative function for the enzyme that aligns its PEP to pyruvate-forming reaction with biosynthetic processes that are specific to early cereal seed development.
Chastain
Chris J
CJ
Department of Biosciences, Minnesota State University-Moorhead, Moorhead, MN 56563, USA. chastain@mnstate.edu
Heck
Jarrod W
JW
Colquhoun
Thomas A
TA
Voge
Dylan G
DG
Gu
Xing-You
XY
D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D018525:Germination / Q000502:physiology; D015151:Immunoblotting; D007527:Isoenzymes; D019684:Magnoliopsida / Q000201:enzymology; D012275:Oryza / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011772:Pyruvate, Orthophosphate Dikinase / Q000235:genetics / Q000378:metabolism*; D012639:Seeds / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
D007527:Isoenzymes; D011772:Pyruvate, Orthophosphate Dikinase