The Triose phosphate-3-phosphoglycerate-phosphate Translocator From Spinach Chloroplasts: Nucleotide Sequence of a Full-Length cDNA Clone and Import of the in Vitro Synthesized Precursor Protein Into Chloroplasts

EMBO J. 1989 Jan;8(1):39-46.

Abstract

The nucleotide sequence of several cDNA clones coding for the phosphate translocator from spinach chloroplasts has been determined. The cDNA clones were selected from a lambda gt10 library prepared from poly(A)+ mRNA of spinach leaves using oligonucleotide probes modeled from amino acid sequences of tryptic peptides prepared from the isolated translocator protein. A 1439 bp insert of one of the clones codes for the entire 404 amino acid residues of the precursor protein corresponding to a mol. wt of 44,234. The full-length clone includes 21 bp at the transcribed non-coding 5' region with the ribosome initiation sequence ACAATGG, a 1212 bp coding region and 199 bp at the non-coding 3' region excluding the poly(A) tail which starts 17 bp downstream from a putative polyadenylation signal, AATAAT. According to secondary structure predictions the mature part of the chloroplast phosphate translocator exhibits high hydrophobicity and consists of at least seven membrane-spanning segments. Using plasmid-programmed wheat germ lysate the precursor protein was synthesized in vitro and could be imported into spinach chloroplasts where it is inserted into the inner envelope membrane.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Chloroplast Proteins
  • Chloroplasts / metabolism*
  • DNA / genetics
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism
  • Membrane Transport Proteins*
  • Molecular Sequence Data
  • Phosphates / metabolism
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Plants / genetics*
  • Plants / metabolism
  • Protein Precursors / metabolism
  • Protein Processing, Post-Translational

Substances

  • Chloroplast Proteins
  • Membrane Proteins
  • Membrane Transport Proteins
  • Phosphates
  • Plant Proteins
  • Protein Precursors
  • DNA

Associated data

  • GENBANK/X13754