Nucleotide sequence of cDNAs encoding the entire precursor polypeptide for thioredoxin m from spinach chloroplasts.
Using the expression vector lambda gt11 and immunochemical detection, six cDNA clones that encode the entire precursor polypeptides for spinach thioredoxin m were isolated and characterized. The ca. 1.0 kb cDNA sequence of the largest clone hybridizes to an RNA species of 1.1 kb. In each instance the cDNA sequences display single open reading frames encoding polypeptides of 181 amino acid residues corresponding to a molecular mass of 19.8 kDa. The sequences of the independently selected cDNAs fall into two classes that are indicative of at least two (closely related) genes for this protein. The amino acid sequences deduced from the cDNA sequences differ to some extent from the amino acid sequence published for spinach thioredoxin m. The sequences predict identical mature proteins of 112-114 amino acids corresponding to a polypeptide molecular mass of ca. 12.4-12.6 kDa, and include stroma-targeting N-terminal transit peptides of 67 residues which are removed during or after import into the organelle. Precursor protein was made in vitro from each of the different cDNA clones and imported into isolated intact chloroplasts. Independent of the cDNA clone used, two isoforms were detected in the chloroplasts after import in each instance. They comigrated with authentic thioredoxin mb and mc. These results indicate that the size variants observed for this protein in vivo result from post-translational modification and do not originate in different genes.
Wedel
N
N
Botanisches Institut, Ludwig-Maximilians-Universität, München, Germany.
Clausmeyer
S
S
Herrmann
R G
RG
Gardet-Salvi
L
L
Schürmann
P
P
D000595:Amino Acid Sequence; D001483:Base Sequence; D054479:Chloroplast Thioredoxins; D002736:Chloroplasts / Q000502:physiology*; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010944:Plants / Q000235:genetics*; D011498:Protein Precursors / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D013879:Thioredoxins / Q000235:genetics*
D054479:Chloroplast Thioredoxins; D011498:Protein Precursors; D013879:Thioredoxins; D004247:DNA