Modulation of glutamine synthetase gene expression in tobacco by the introduction of an alfalfa glutamine synthetase gene in sense and antisense orientation: molecular and biochemical analysis.
A glutamine synthetase (GS) cDNA isolated from an alfalfa cell culture cDNA library was found to represent a cytoplasmic GS. The full-length alfalfa GS1 coding sequence, in both sense and antisense orientation and under the transcriptional control of the cauliflower mosaic virus 35S promoter, was introduced into tobacco. Leaves of tobacco plants transformed with the sense construct contained greatly elevated levels of GS transcript and GS polypeptide which assembled into active enzyme. Leaves of the plants transformed with the antisense GS1 construct showed a significant decrease in the level of both GS1 and GS2 polypeptides and GS activity, but did not show any significant decrease in the level of endogenous GS mRNA. We have proposed that antisense inhibition using a heterologous antisense GS RNA occurs at the level of translation. Our results also suggest that the post-translational assembly of GS subunits into a holoenzyme requires an additional factor(s) and is under regulatory control.
Temple
S J
SJ
Plant Genetic Engineering Labs/Department of Agronomy and Horticulture, New Mexico State University, Las Cruces 88003.
Knight
T J
TJ
Unkefer
P J
PJ
Sengupta-Gopalan
C
C
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis; D002478:Cells, Cultured; D003001:Cloning, Molecular; D015536:Down-Regulation; D015870:Gene Expression; D014644:Genetic Variation; D005974:Glutamate-Ammonia Ligase / Q000096:biosynthesis / Q000235:genetics*; D050537:Hydroxypyruvate Reductase; D000455:Medicago sativa / Q000201:enzymology / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D010730:Phosphoenolpyruvate Carboxylase / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics*; D010947:Plants, Toxic; D011487:Protein Conformation; D016372:RNA, Antisense / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D013045:Species Specificity; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics*; D014158:Transcription, Genetic; D014170:Transformation, Genetic
D010940:Plant Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D000429:Alcohol Oxidoreductases; D050537:Hydroxypyruvate Reductase; D010730:Phosphoenolpyruvate Carboxylase; D005974:Glutamate-Ammonia Ligase