Identification and organization of chloroplastic and cytosolic L-myo-inositol 1-phosphate synthase coding gene(s) in Oryza sativa: comparison with the wild halophytic rice, Porteresia coarctata

Sudipta Ray, Barunava Patra, Aparajita Das-Chatterjee, Arnab Ganguli, Arun Lahiri Majumder

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

The gene coding for rice chloroplastic L-myo-inositol-1-phosphate synthase (MIPS; EC 5.5.1.4) has been identified by matrix-assisted laser desorption time-of-flight mass spectrometry analysis of the purified and immunologically cross-reactive approximately 60 kDa chloroplastic protein following two-dimensional polyacrylamide gel electrophoresis, which exhibited sequence identity with the cytosolic MIPS coded by OsINO1-1 gene. A possible chloroplastic transit peptide sequence was identified upstream of the OsINO1-1 gene upon analysis of rice genome. RT-PCR and confocal microscope studies confirmed transcription, effective translation and its functioning as a chloroplast transit peptide. Bioinformatic analysis mapped the chloroplastic MIPS (OsINO1-1) gene on chromosome 3, and a second MIPS gene (OsINO1-2) on chromosome 10 which lacks conventional chloroplast transit peptide sequence as in OsINO1-1. Two new PcINO1 genes, with characteristic promoter activity and upstream cis-elements were identified and cloned, but whether these proteins can be translocated to the chloroplast or not is yet to be ascertained. Electrophoretic mobility shift assay carried out with nuclear extract of Porteresia coarctata leaves grown under both control and stressed condition shows binding of nuclear proteins with the upstream elements. Nucleotide divergence among the different Oryza and Porteresia INO1 genes were calculated and compared.

Original languageEnglish
Pages (from-to)1211-27
Number of pages17
JournalPlanta
Volume231
Issue number5
DOIs
StatePublished - Apr 2010

Keywords

  • Amino Acid Sequence
  • Base Sequence
  • Chloroplast Proteins
  • Chloroplasts/enzymology
  • Codon, Terminator/genetics
  • Cytosol/enzymology
  • Databases, Genetic
  • Electrophoretic Mobility Shift Assay
  • Evolution, Molecular
  • Genes, Plant/genetics
  • Glucuronidase/metabolism
  • Green Fluorescent Proteins/metabolism
  • Introns/genetics
  • Molecular Sequence Data
  • Myo-Inositol-1-Phosphate Synthase/chemistry
  • Open Reading Frames/genetics
  • Oryza/enzymology
  • Phylogeny
  • Protein Sorting Signals/genetics
  • Proteomics
  • Regulatory Sequences, Nucleic Acid/genetics
  • Ribosomes/metabolism
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

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