Identification of a Ribose-Phosphate Pyrophosphokinase that Can Interact In Vivo with the Anaphase Promoting Complex/Cyclosome

Int J Mol Sci. 2017 Mar 30;18(4):617. doi: 10.3390/ijms18040617.

Abstract

5-Phospho-d-ribosyl-1-diphosphate (PRPP) synthase (PRS) catalyzes the biosynthesis of PRPP, which is an important compound of metabolism in most organisms. However, no PRS genes have been cloned, let alone studied for their biological function in rubber tree. In this study, we identify a novel protein (PRS4) that interacts in vivo with rubber tree anaphase promoting complex/cyclosome (APC/C) subunit 10 (HbAPC10) by yeast two-hybrid assays. PRS4 has been cloned from rubber tree and named as HbPRS4. Blastp search in the genome of Arabidopsis thaliana showed that HbPRS4 shared the highest similarity with AtPRS4, with 80.71% identity. qRT-PCR was used to determine the expression of HbPRS4 in different tissues and under various treatments. HbPRS4 was preferentially expressed in the bark. Moreover, the expression level of HbPRS4 was significantly induced by the proteasome inhibitor MG132 treatment, suggesting it might be regulated by the ubiquitin/26S proteasome pathway. The amount of HbPRS4 transcript was obviously decreased after mechanical wounding and abscisic acid (ABA) treatments, while a slight increase was observed at 24 h after ABA treatment. HbPRS4 transcript in the latex was significantly upregulated by ethephon (ET) and methyl jasmonate (MeJA) treatments. These results suggested that HbPRS4 may be a specific substrate of HbAPC10 indirectly regulating natural rubber biosynthesis in rubber tree.

Keywords: ">d-ribosyl-1-diphosphate; 5-phospho-; anaphase promoting complex/cyclosome; ribose-phosphate pyrophosphokinase; rubber tree.

MeSH terms

  • Abscisic Acid / pharmacology
  • Acetates / pharmacology
  • Anaphase-Promoting Complex-Cyclosome / metabolism*
  • Cyclopentanes / pharmacology
  • Gene Expression Regulation, Plant / drug effects
  • Hevea / genetics
  • Hevea / metabolism
  • Leupeptins / pharmacology
  • Oxylipins / pharmacology
  • Plant Bark / metabolism
  • Plant Proteins / chemistry
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*
  • Protein Binding
  • Ribose-Phosphate Pyrophosphokinase / chemistry
  • Ribose-Phosphate Pyrophosphokinase / genetics
  • Ribose-Phosphate Pyrophosphokinase / metabolism*

Substances

  • Acetates
  • Cyclopentanes
  • Leupeptins
  • Oxylipins
  • Plant Proteins
  • Abscisic Acid
  • methyl jasmonate
  • Anaphase-Promoting Complex-Cyclosome
  • Ribose-Phosphate Pyrophosphokinase
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde