Eight small subunits of Euglena ribulose 1-5 bisphosphate carboxylase/oxygenase are translated from a large mRNA as a polyprotein.
The small subunit (SSU) of ribulose 1-5 bisphosphate carboxylase/oxygenase is a 15 kd protein in Euglena gracilis. The protein is synthesized as a 130 kd precursor as shown by immunoprecipitation of in vitro translation products and confirmed by immunoprecipitation of in vivo pulse-labeled Euglena proteins. From the published SSU amino acid sequence, an oligonucleotide was synthesized that specifically hybridizes to a large mRNA whose length (approximately 4.3 kb) is consistent with the precursor size. The complete nucleotide sequence of the SSU mRNA was obtained by sequencing a cDNA clone from a lambda gt11 library and completed by direct mRNA sequencing. We report for the first time the complete sequence of a large mRNA and show that it encodes eight consecutive SSU mature molecules. The deduced precursor amino acid sequence shows that the amino terminus of the first SSU molecule is preceded by a 134 amino acid peptide which is cleaved during the maturation process. This long transit peptide exhibits features characteristic of signal peptides involved in the secretion of proteins through the endoplasmic reticulum. This is in agreement with the idea that the third (outer) membrane of the Euglena chloroplast envelope is of endoplasmic reticulum origin.
Chan
R L
RL
Institut de Biologie Moléculaire des Plantes du CNRS, Université Louis Pasteur, Strasbourg, France.
Keller
M
M
Canaday
J
J
Weil
J H
JH
Imbault
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002736:Chloroplasts / Q000201:enzymology; D004247:DNA / Q000235:genetics; D004792:Enzyme Precursors / Q000235:genetics; D005056:Euglena gracilis / Q000201:enzymology / Q000235:genetics*; D015723:Gene Library; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000235:genetics*; D015183:Restriction Mapping; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics*
D004792:Enzyme Precursors; D046911:Macromolecular Substances; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D004247:DNA; D012273:Ribulose-Bisphosphate Carboxylase