Cloning rbpl-1: C. elegans ortholog of RBBP6, activating factor of the RNA cleavage and polyadenylation complex
The RNA Cleavage and Polyadenylation Complex (CPC) is responsible for cleaving and preparing pre-mRNA through the addition of a poly(A) tail; this long strand of adenosines aids in stabilizing the pre-mRNA before it is translated into amino acids to make protein. However, the CPC's mechanisms, each with vastly different functions and complexities, are yet to be fully understood. The CPC is highly conserved among different organisms, allowing us to study human CPC using more accessible models such as C. elegans. Because of this, we can investigate the components of the C. elegansCPC in order to gain a deeper understanding of the mechanisms underlying this fundamental process in humans. Our goal for this project was to clone rbpl-1, the gene encoding for the worm ortholog of the human protein RBBP6, whose role in cleavage and polyadenylation has only recently been discovered. Although it is not considered a component of the CPC, RBBP6 activates an endonuclease within the CPC, CPSF73, which is responsible for cleaving pre-mRNA. Cloning rbpl-1will produce a resource that can be used in future experimentation on its role in cleavage and polyadenylation. Currently, we are optimizing the conditions necessary to produce rbpl-1 cDNA from total RNA to clone into an entry vector.