Molecular characterization of a cDNA clone encoding glutamine synthetase from a gymnosperm, Pinus sylvestris.
A full-length cDNA clone (pGSP114) encoding glutamine synthetase was isolated from a lambda gt11 library of the gymnosperm Pinus sylvestris. Nucleotide sequence analysis showed that pGSP114 contains an open reading frame encoding a protein of 357 amino acid residues with a calculated molecular mass of 39.5 kDa. The derived amino acid sequence was more homologous to cytosolic (GS1) (78-82%) than to chloroplastic (GS2) (71-75%) glutamine synthetase in angiosperms. The lack of N-terminal presequence and C-terminal extension which define the primary structure of GS2, also supports that the isolated cDNA encodes cytosolic GS. Southern blot analysis of genomic DNA from P. sylvestris and P. pinaster suggests that GS may be encoded by a small gene family in pine. GS mRNA was more abundant in cotyledons and stems than in roots of both Scots and maritime pines. Western blot analysis in P. sylvestris seedlings showed that only one GS polypeptide, similar in size to GS1 in P. pinaster, could be detected in several different tissues. Our results suggest that cytosolic GS is mainly responsible for glutamine biosynthesis in pine seedlings.
Cantón
F R
FR
Laboratorio de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Málaga, Spain.
García-Gutiérrez
A
A
Gallardo
F
F
de Vicente
A
A
Cánovas
F M
FM
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA; D005974:Glutamate-Ammonia Ligase / Q000235:genetics*; D008969:Molecular Sequence Data; D041605:Pinus sylvestris; D010944:Plants / Q000201:enzymology / Q000235:genetics*; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic
D004247:DNA; D005974:Glutamate-Ammonia Ligase