Department of Plant Biology, University of Illinois, 1201 W. Gregory Drive, Urbana, Illinois 61801.
Plant Physiol. 1992 Oct;100(2):812-9. doi: 10.1104/pp.100.2.812.
The expression of calmodulin mRNA and protein were measured during a growth cycle of carrot (Daucus carota L.) cells grown in suspension culture. A full-length carrot calmodulin cDNA clone isolated from a lambdagt10 library was used to measure steady-state calmodulin mRNA levels. During the exponential phase of culture growth when mitotic activity and oxidative respiration rates were maximal, calmodulin mRNA levels were 4- to 5-fold higher than they were during the later stages of culture growth, when respiration rates were lower and growth was primarily by cell expansion. Net calmodulin polypeptide synthesis, as measured by pulse-labeling in vivo with [(35)S]methionine, paralleled the changes in calmodulin steady-state mRNA level during culture growth. As a consequence, net calmodulin polypeptide synthesis declined 5- to 10-fold during the later stages of culture growth. The qualitative spectrum of polypeptides synthesized and accumulated by the carrot cells during the course of a culture cycle, however, remained largely unchanged. Calmodulin polypeptide levels, in contrast to its net synthesis, remained relatively constant during the exponential phases of the culture growth cycle and increased during the later stages of culture growth. Our data are consistent with increased calmodulin polypeptide turnover associated with periods of rapid cell proliferation and high levels of respiration.
在悬浮培养的胡萝卜(Daucus carota L.)细胞的生长周期中,测量了钙调蛋白 mRNA 和蛋白质的表达。从 lambdagt10 文库中分离出的全长胡萝卜钙调蛋白 cDNA 克隆被用于测量稳态钙调蛋白 mRNA 水平。在培养生长的指数期,当有丝分裂活性和氧化呼吸速率达到最大值时,钙调蛋白 mRNA 水平比培养生长后期高 4 到 5 倍,此时呼吸速率较低,生长主要通过细胞扩张。通过体内用 [(35)S]甲硫氨酸脉冲标记测量净钙调蛋白多肽合成,与培养生长过程中钙调蛋白稳态 mRNA 水平的变化平行。因此,在培养生长后期,净钙调蛋白多肽合成下降了 5 到 10 倍。然而,在培养周期过程中,胡萝卜细胞合成和积累的多肽的定性谱基本保持不变。与净合成相反,钙调蛋白多肽水平在培养生长周期的指数期相对保持稳定,并在培养生长后期增加。我们的数据与与快速细胞增殖和高呼吸水平相关的钙调蛋白多肽周转率增加一致。