Chloroplast and cytosolic glutamine synthetase are encoded by homologous nuclear genes which are differentially expressed in vivo

• Published in: The Journal of biological chemistry 1988-07, Vol.263 (20), p.9651-9657
• Main Authors: Tingey, S V, Tsai, F Y, Edwards, J W, Walker, E L, Coruzzi, G M
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Base Sequence; Cell Nucleus - metabolism; Chloroplasts - enzymology; Cytosol - enzymology; DNA - genetics; DNA, Recombinant - metabolism; Electrophoresis, Polyacrylamide Gel; Fabaceae; Gene Expression Regulation; Glutamate-Ammonia Ligase - genetics; Immunoassay; Light; Molecular Sequence Data; Pisum sativum; Plants, Medicinal; Protein Biosynthesis; Protein Precursors - genetics; RNA, Messenger - biosynthesis; RNA, Messenger - radiation effects; Sequence Homology, Nucleic Acid
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)81566-1

We have shown that the individual members of the plant gene family for glutamine synthetase (GS) are differentially expressed in vivo, and each encode distinct GS polypeptides which are targeted to different subcellular compartments (chloroplast or cytosol). At the polypeptide level, chloroplast GS (GS2) and cytosolic GS (GS1 and GSn) are distinct and show an organ-specific distribution. We have characterized full length cDNA clones encoding chloroplast or cytosolic GS of pea. In vitro translation products encoded by three different GS cDNA clones, correspond to the mature GS2, GS1, and GSn polypeptides present in vivo. pGS185 encodes a precursor to the chloroplast GS2 polypeptide as shown by in vitro chloroplast uptake experiments. The pGS185 translation product is imported into the chloroplast stroma and processed to a polypeptide which corresponds in size and charge to that of mature chloroplast stromal GS2 (44 kDa). The 49 amino terminal amino acids encoded by pGS185 are designated as a chloroplast transit peptide by functionality in vitro, and amino acid homology to other transit peptides. The cytosolic forms of GS (GS1 and GSn) are encoded by highly homologous but distinct mRNAs. pGS299 encodes the cytosolic GS1 polypeptide (38 kDa), while pGS341 (Tingey, S. V., Walker, E. L., and Coruzzi, G. M. (1987) EMBO. J. 6, 1-9) encodes a cytosolic GSn polypeptide (37 kDa). The homologous nuclear genes for chloroplast and cytosolic GS show different patterns of expression in vivo. GS2 expression in leaves is modulated by light, at the level of steady state mRNA and protein, while the expression of cytosolic GS is unaffected by light. The light-induced expression of GS2 is due at least in part to a phytochrome mediated response. Nucleotide sequence analysis indicates that chloroplast and cytosolic GS have evolved from a common ancestor and suggest a molecular mechanism for chloroplast evolution.