Evidence for the translational control of storage protein gene expression in oat seeds.
We employed a rapid fractionation method coupled with a sensitive enzyme-linked immunosorbent assay to quantify the globulins and avenins in developing and mature oat seeds. On a molar basis, there is approximately 10-11 times as much globulin as avenin. Pulse labeling of endosperm proteins indicated that the rate of globulin synthesis is approximately nine times that of avenin. In addition, neither protein class showed any signs of degradation during this experiment. Analysis of the storage protein mRNAs indicates that both globulin and avenin transcripts are associated with membrane-bound polysomes and are found in similar concentrations within the membrane-bound polysome fraction. We found that avenin and globulin mRNAs are fully loaded with ribosomes, suggesting that initiation is not rate-limiting for translation of either protein. Rates of globulin and avenin synthesis were similar when synthetic storage protein mRNAs were translated in vitro. Translation of equimolar amounts of globulin and avenin mRNAs in the same reaction showed equivalent amounts of protein synthesized when compared with globulin and avenin mRNAs translated in separate reaction mixes. We propose that translation elongation or termination reactions are likely regulatory steps for controlling storage protein synthesis in oat endosperm.
Boyer
S K
SK
Department of Plant Sciences, University of Arizona, Tucson 85721.
Shotwell
M A
MA
Larkins
B A
BA
D001345:Autoradiography; D002523:Edible Grain / Q000235:genetics* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation; D017343:Genes, Plant; D005916:Globulins / Q000235:genetics; D010940:Plant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D055315:Prolamins; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D012639:Seeds / Q000378:metabolism; D013431:Sulfates / Q000378:metabolism; D014158:Transcription, Genetic
D005916:Globulins; D010940:Plant Proteins; D055315:Prolamins; D012333:RNA, Messenger; D013431:Sulfates