Abstract
Abstract: :
Purpose: Different types of regulation are utilized to produce a robust circadian clock, including regulation at the transcriptional, post-transcriptional, and translational levels. A screen for rhythmic messages that may be involved in such circadian control identified nocturnin, a novel gene that displays high amplitude circadian expression in the Xenopus laevis retina, with peak mRNA levels in the early night. Expression of nocturnin mRNA is confined to the clock-containing photoreceptor cell layer within the retina. Methods: Deadenylase activity was measured in vitro from recombinant GST-nocturnin fusion protein. Protein localization and temporal accumulation were measured by immunohistochemistry and western blot analyses. Results: In these studies we show that nocturnin removes the poly(A) tail from a synthetic RNA substrate in a process known as deadenylation. Nocturnin nuclease activity is magnesium-dependent, as addition of EDTA or mutation of the residue predicted to bind magnesium disrupts deadenylation. Substrate preference studies show that nocturnin is an exonuclease that specifically degrades the 3' poly (A) tail. While nocturnin is rhythmically expressed in the cytoplasm of the retinal photoreceptor cells, the only other described vertebrate deadenylase, PARN, is constitutively present in most retinal cells, including the photoreceptors. Conclusions: The distinct spatial and temporal expression of nocturnin and PARN suggests there may be specific mRNA targets of each deadenylase. Since deadenylation regulates mRNA decay and/or translational silencing, we propose that nocturnin deadenylates clock-related transcripts in a novel mechanism for post-transcriptional regulation in the circadian clock or its outputs.
Keywords: photoreceptors • circadian rhythms • gene/expression