Regulation of a specific circadian clock output pathway by lark, a putative RNA-binding protein with repressor activity

Abstract

An endogenous clock within the Drosophila brain regulates circadian rhythms in adult eclosion and locomotor activity. Although molecular elements of the Drosophila circadian clock have been well characterized, little is known about the clock output pathways that mediate the control of rhythmic events. Previous genetic analysis indicates that a gene known as lark encodes an element of the clock output pathway regulating adult eclosion. We now present evidence that lark encodes a novel member of the RNA recognition motif (RRM) class of RNA-binding proteins. Similar to other members of this protein superfamily, lark contains two copies of a bipartite consensus RNA-binding motif. Unlike any other RRM family member, however, lark protein also contains a distinct class of nucleic acid binding motif, a retroviral-type zinc finger, that is present in the nucleocapsid protein of retroviruses and in several eukaryotic proteins. In contrast to identified clock elements, lark mRNA does not exhibit diurnal fluctuations in abundance in late pupae or in adult heads. Thus rhythmic transcription of the gene does not contribute to the temporal regulation of eclosion by lark protein. Gene dosage experiments show that decreased or increased lark product, respectively, leads to an early or late eclosion phenotype, indicating that the protein negatively regulates the eclosion process. It is postulated that lark is required for the posttranscriptional repression of genes encoding other elements of this clock output pathway.