Bell-Pedersen D, Shinohara M L, Loros J J, Dunlap J C
Department of Biochemistry, Darmouth Medical School, Hanover, NH 03755, USA.
Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):13096-101. doi: 10.1073/pnas.93.23.13096.
An endogenous circadian biological clock controls the temporal aspects of life in most organisms, including rhythmic control of genes involved in clock output pathways. In the fungus Neurospora crassa, one pathway known to be under control of the clock is asexual spore (conidia) development. To understand more fully the processes that are regulated by the N. crassa circadian clock, systematic screens were carried out for genes that oscillate at the transcriptional level. Time-of-day-specific cDNA libraries were generated and used in differential screens to identify six new clock-controlled genes (ccgs). Transcripts specific for each of the ccgs preferentially accumulate during the late night to early morning, although they vary with respect to steady-state mRNA levels and amplitude of the rhythm. Sequencing of the ends of the new ccg cDNAs revealed that ccg-12 is identical to N. crassa cmt encoding copper metallothionein, providing the suggestion that not all clock-regulated genes in N. crassa are specifically involved in the development of conidia. This was supported by finding that half of the new ccgs, including cmt(ccg-12), are not transcriptionally induced by developmental or light signals. These data suggest a major role for the clock in the regulation of biological processes distinct from development.
内源性昼夜生物钟控制着大多数生物体生命活动的时间节律,包括对生物钟输出途径中相关基因的节律性调控。在粗糙脉孢菌中,已知受生物钟调控的一条途径是无性孢子(分生孢子)的发育。为了更全面地了解受粗糙脉孢菌昼夜生物钟调控的过程,我们针对在转录水平上振荡的基因进行了系统筛选。构建了特定时间点的cDNA文库,并用于差异筛选,以鉴定出六个新的生物钟调控基因(ccgs)。尽管每个ccg的转录本在稳态mRNA水平和节律幅度上有所不同,但它们都优先在深夜至清晨积累。对新ccg cDNA末端的测序显示,ccg - 12与编码铜金属硫蛋白的粗糙脉孢菌cmt相同,这表明粗糙脉孢菌中并非所有受生物钟调控的基因都专门参与分生孢子的发育。这一观点得到了如下发现的支持:包括cmt(ccg - 12)在内的一半新ccg,在发育或光信号刺激下不会发生转录诱导。这些数据表明,生物钟在调控与发育不同的生物学过程中发挥着重要作用。