Sequencing and Analysis of Centrosomal RNAs

Marine Biological Laboratory, Woods Hole MA

Investigators

Abstract

Intellectual Merit The centrosome in animal cells is composed of a pair of centrioles surrounded by a relatively ill-defined pericentriolar matrix. Centrosomes serve as the major microtubule organizing center in animal cells, carry the mother centriole which is the basal body progenitor, and organize the assembly of the mitotic spindle during cell division. Centrosomes duplicate only once per cell cyle to ensure development of a normal bipolar spindle. The initial event in centrosome duplication is centriole replication, which is generative, semiconservative, and independent of the nucleus. These and other observations have led investigators to propose that centrosomes may contain their own complement of nucleic acids, possibly representative of an organellar genome as has been described for mitochondria and chloroplasts. Until recently, the only consensus developed during the past 50 years is that centrosomes do not contain DNA. The existence of centrosomal RNA (cnRNA) remained an open possibility. Research in this investigator's laboratory and others has now resolved this decades-old question in the affirmative, although the source and function of cnRNAs remain unknown and controversial. In this project, high throughput sequencing technology and informatics will be used to characterize cnRNAs from the surf clam, Spisula solidissima, and identify related molecules in other species. This will open the way for future phylogenetic and functional analyses to help determine the evolutionary origin and physiological roles of cnRNAs. Broader Impact This project will help provide training for one postdoctoral scientist and one graduate student at the Marine Biological Laboratory. Opportunities for undergraduate training are also available in the PI's laboratory. In addition to submitting raw sequence data to NCBI, a database for centrosomal RNAs will be made available to other investigators via web server. The database will be hosted on high performance servers at the MBL that are already in use for other, similar projects. Finally, should the results of this study uphold hypotheses for an exogenous origin for the centriole-centrosome system, it will represent a significant contribution to our understanding of the role played by symbiogenesis in the origin and evolution of the eukaryotes.