Interplay of CDKs and cyclins with glycolytic regulatory enzymes PFK and PK

• Published in: Journal of plant physiology 2024-12, Vol.303, p.154378, Article 154378
• Main Authors: Lara-Núñez, Aurora, Guerrero-Molina, Estefany Damaris, Vargas-Cortez, Teresa, Vázquez-Ramos, Jorge Manuel
• Format: Article
• Language: English
• Subjects: Cyclin-dependent kinase; Cyclin-Dependent Kinases - genetics; Cyclin-Dependent Kinases - metabolism; Cyclins - metabolism; Germination; Glycolysis; Maize; PFK; Phosphorylation; Plant Proteins - genetics; Plant Proteins - metabolism; Pyruvate Kinase - metabolism; Zea mays - enzymology; Zea mays - genetics; Zea mays - metabolism
• ISSN: 0176-1617; 1618-1328; 1618-1328
• DOI: 10.1016/j.jplph.2024.154378

In plants, as in all eukaryotes, the cell cycle is regulated by the heterodimer formed by cyclins (Cycs) and cyclin-dependent kinases (CDKs), that phosphorylate serine/threonine residues in target proteins. The extensive involvement of these heterodimers in nuclear cell cycle-related processes has been demonstrated. However, recent findings have linked Cyc-CDK complexes to the regulation of cytosolic processes, including various metabolic pathways, suggesting close coordination between the cell cycle and catabolic/anabolic processes to maintain cellular energy homeostasis. This study extends the analysis of Cyc-CDK complex regulation in maize to two key regulators of glycolysis: phosphofructose kinase (PFK) and pyruvate kinase (PK). Both are cytosolic enzymes, highly regulated positively and negatively by different metabolites, showing a similar activation pattern in their homotetrameric form and low activity when as dimers/monomers. Each enzyme exhibits two putative minimal phosphorylation motives for Cyc-CDKs, conserved in some plant species and in four (PFK) and three (PK) isoforms in maize. This work demonstrates that both enzymes are active with fluctuating levels of activity along maize germination; also, that they associate with different maize Cycs and CDKs as demonstrated by pull-down assays, as well as their in vitro phosphorylation by recombinant CycD;2-CDKA or CycD2;2-CDKB complexes. Additionally, the inhibition of PFK and PK activity following phosphorylation by active Cycs-CDKB complexes obtained by immunoprecipitation from imbibed embryonic axis protein extracts suggests a narrow and negative regulation of glycolysis as the cell cycle progresses. A decreased carbon flow through this pathway is proposed to divert carbon from sugars towards the oxidative pentose phosphate pathway, thereby promoting de novo nucleic acid synthesis precursors to stimulate cell cycle progression. [Display omitted] Maize PFK and PK isoforms contain two CDK phosphorylation motifs.PFK and PK interacted distinctively with members of the Cyclin/CDK complexes in maizePFK and PK were phosphorylated in vitro by recombinant CycD2;2/CDK (A/B)The activities of PFK and PK were inhibited following phosphorylation by Cyc/CDKB