Evans J H, Keller L R
Department of Biological Science, The Florida State University, Tallahassee 32306-3050, USA.
Plant Mol Biol. 1997 Feb;33(3):467-81. doi: 10.1023/a:1005727806897.
Acid shock of Chlamydomonas results in flagellar excision and induction of flagellar protein RNAs. The magnitude of flagellar RNA accumulations after flagellar excision by mechanical shear depends on the extracellular Ca2+ concentration. In this report, we demonstrate that the magnitude and duration of flagellar RNA accumulations are signaled by an acid shock-induced Ca2+ influx. RNA accumulations were greater in cells acid shocked in 500 microM CaCl2 than in 200 microM CaCl2, although the accumulation durations were similar. RNA accumulations of lower magnitude and shorter duration were observed in cells in Ca2+-containing buffer treated with CdCl2. RNA accumulations were of still lower magnitude and shorter duration in cells shocked in buffer without added CaCl2 than in cells shocked in 200 or 500 microM CaCl2 or in the presence of CdCl2. RNA accumulations similar to those in cells shocked in buffer without added CaCl2 were measured in cells following acid shock in buffer containing 200 microM CaCl2 and supplemented with neomycin, ruthenium red, or LaCl3. Acid shock of the adf-1 mutant resulted in RNA accumulations of shorter duration and lower magnitude than those measured in adf-1 cells stimulated by mechanical shear. These results are consistent with an hypothesis that acid shock generates two genetically and pharmacologically distinct signals governing flagellar RNA induction; the first signal is independent of a Ca2+ influx and flagellar excision and results in low magnitude accumulations of short duration, and the second is a consequence of a Ca2+ influx and results in accumulations of high magnitude and long duration.
衣藻的酸休克会导致鞭毛切除并诱导鞭毛蛋白RNA的产生。通过机械剪切切除鞭毛后,鞭毛RNA积累的程度取决于细胞外Ca2+浓度。在本报告中,我们证明了鞭毛RNA积累的程度和持续时间是由酸休克诱导的Ca2+内流发出信号的。尽管积累持续时间相似,但在500微摩尔氯化钙中酸休克处理的细胞中,RNA积累比在200微摩尔氯化钙中酸休克处理的细胞中更大。在用氯化镉处理的含Ca2+缓冲液中的细胞中,观察到RNA积累的程度较低且持续时间较短。与在200或500微摩尔氯化钙中酸休克处理的细胞或在存在氯化镉的情况下酸休克处理的细胞相比,在未添加CaCl2的缓冲液中酸休克处理的细胞中,RNA积累的程度更低且持续时间更短。在含有200微摩尔氯化钙并添加新霉素、钌红或氯化镧的缓冲液中酸休克处理的细胞中,测量到的RNA积累与在未添加CaCl2的缓冲液中酸休克处理的细胞中的RNA积累相似。adf-1突变体的酸休克导致RNA积累的持续时间比机械剪切刺激的adf-1细胞中测量到的持续时间短,程度也更低。这些结果与一个假设一致,即酸休克产生两个在遗传和药理学上不同的信号来控制鞭毛RNA的诱导;第一个信号独立于Ca2+内流和鞭毛切除,导致短时间低程度的积累,第二个信号是Ca2+内流的结果,导致高程度和长时间的积累。