Multiple plant RNA binding proteins identified by PCR: expression of cDNAs encoding RNA binding proteins targeted to chloroplasts in Nicotiana plumbaginifolia

Mol Gen Genet. 1992 Sep;234(3):390-400. doi: 10.1007/BF00538698.

Abstract

Pre-mRNA processing in eukaryotic cells requires the participation of multiple protein factors and ribonucleoprotein particles. One class of proteins involved in this process are RNA-binding proteins, which contain a domain of ca. 90 amino acids with a characteristic ribonucleoprotein consensus sequence (RNP-CS). A PCR approach that is suitable for the characterization of RNP-CS-type proteins is described. Fifteen different RNA-binding domains were amplified from Nicotiana tabacum (tobacco) using oligonucleotide primers specific for the sequences (K/R)G(F/Y)(G/A)FVX(F/Y) and (L/I/V)(F/Y)(V/I)(G/K)(N/G)L, which are conserved in known RNP-CS proteins. Using the tobacco domains as probes, cDNAs encoding two RNA-binding proteins, each containing two RNP-CS-type domains, were characterized in N. plumbaginifolia. The proteins, designated CP-RBP30 and CP-RBP31, are targeted to chloroplasts as demonstrated by expression of epitope-tagged cDNAs in transfected protoplasts, followed by indirect immunofluorescence. High levels of mRNA for each protein were found in leaves but not in roots, and expression of the CP-RBP31 mRNA was strongly regulated by light. The N. plumbaginifolia proteins described in this work are distinct from chloroplast RNA-binding proteins characterized recently in tobacco and spinach.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Chloroplasts / physiology*
  • Consensus Sequence
  • DNA / genetics
  • Fluorescent Antibody Technique
  • Gene Expression
  • Light
  • Molecular Sequence Data
  • Nicotiana / genetics*
  • Oligodeoxyribonucleotides / chemistry
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Plants, Toxic*
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics
  • RNA-Binding Proteins / genetics*
  • RNA-Binding Proteins / metabolism
  • Ribonucleoproteins / chemistry
  • Ribonucleoproteins / genetics
  • Ribonucleoproteins / metabolism
  • Sequence Alignment

Substances

  • Oligodeoxyribonucleotides
  • Plant Proteins
  • RNA, Messenger
  • RNA-Binding Proteins
  • Ribonucleoproteins
  • DNA

Associated data

  • GENBANK/X65117
  • GENBANK/X65118