Summary Cellulose is an essential component of plant cell walls and an economically important source of food, paper, textiles, and biofuel. Despite its economic and biological significance, the regulation of cellulose biosynthesis is poorly understood. Phosphorylation and dephosphorylation of cellulose synthases (CESAs) were shown to impact the direction and velocity of cellulose synthase complexes (CSCs). However, the protein kinases that phosphorylate CESAs are largely unknown. We conducted research inArabidopsis thalianato reveal protein kinases that phosphorylate CESAs.In this study, we used yeast two‐hybrid, protein biochemistry, genetics, and live‐cell imaging to reveal the role of calcium‐dependent protein kinase32 (CPK32) in the regulation of cellulose biosynthesis inA. thaliana.We identified CPK32 using CESA3 as a bait in a yeast two‐hybrid assay. We showed that CPK32 phosphorylates CESA3 while it interacts with both CESA1 and CESA3. Overexpressing functionally defective CPK32 variant and phospho‐dead mutation of CESA3 led to decreased motility of CSCs and reduced crystalline cellulose content in etiolated seedlings. Deregulation of CPKs impacted the stability of CSCs.We uncovered a new function of CPKs that regulates cellulose biosynthesis and a novel mechanism by which phosphorylation regulates the stability of CSCs.
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B4 Raf‐like MAPKKK RAF24 regulates Arabidopsis thaliana flowering time through HISTONE MONO ‐ UBIQUITINATION 2
Summary The timing of flowering is a critical agronomic trait governed by an extensive and sophisticated regulatory network. To date, limited understanding of how posttranslational modifications regulate flowering time exists. Here, using Arabidopsis, we resolve a role for the B4 Raf‐like MAPKKK protein kinase RAF24 in regulating flowering time.Loss of RAPIDLY ACCELERATED FIBROSARCOMA24 (RAF24) led to premature flowering time through altered expression ofFLCandFT. Comparative phosphoproteomic analysis ofraf24and wild‐type plants revealed a list of known flowering‐related phosphoproteins from distinct flowering pathways displaying downregulated phosphorylation. Of these, the RING‐type ubiquitin ligase HISTONE MONO‐UBIQUITINATION 2 (HUB2) lacked phosphorylation in the absence of RAF24.Absence of RAF24 induced H2Bub1 overaccumulation, with protein–protein interactome analysis of HUB2 in the presence and absence of RAF24 influencing HUB2 protein interaction partners, such as H2B. HUB2 was also found to physically interact with SUCROSE NONFERMENTING KINASE 2.4 (SnRK2.4) and SnRK2.6, known substrates of RAF24. Using phospho‐mimetic and phospho‐ablative plant lines, we then validated the importance of HUB2 phosphorylation at serine 314 (S314) in maintaining appropriate flowering time.Our findings uncovered a novel biological role of RAF24, as a higher‐order flowering regulator, while further implicating HUB2 as a centerpiece of flowering time regulation.
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- Award ID(s):
- 2420360
- PAR ID:
- 10683172
- Publisher / Repository:
- New Phytologist
- Date Published:
- Journal Name:
- New Phytologist
- Volume:
- 247
- Issue:
- 2
- ISSN:
- 0028-646X
- Page Range / eLocation ID:
- 897 to 915
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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