2011 Rustbelt RNA Meeting
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Poster number 82 submitted by Trinh Tat

Investigating the non-canonical polyadenylation machinery in D. melanogaster S2 cells

Trinh T. Tat (RNA Center - Biochemistry Dept. - Case Western Reserve University), Raul Jobava (RNA Center - Biochemistry Dept. - Case Western Reserve University), Patricia Maroney (RNA Center - Biochemistry Dept. - Case Western Reserve University), Timothy Nilsen (RNA Center - Biochemistry Dept. - Case Western Reserve University)

Abstract:
Cleavage and polyadenylaton at the 3’ end of transcripts are required post-transcriptional modifications contributing to mRNA maturation in eukaryotes. This process is critical for transcription termination, nuclear export, stability and efficient translation of mRNA. It has been known that alternative polyadenylation can alter the coding sequence of the encoded protein and have an effect on how much protein is made from a transcript. By using a sensitive assay for poly(A) tail length developed in our lab followed by sequencing, we found that the Hid and Reaper transcripts in S2 cells do not contain the normal poly(A) tail compared with several controls, such as actin or Par-6. In addition, through sequencing, we found that the sequence immediately downstream of the hexanucleotide polyadenylation signal in this mRNA is the same as the genomic DNA sequence. Therefore, the Hid and Reaper pre-mRNAs are either cleaved but not polyadenylated or deadenylated. We are currently asking what are the essential cis- and trans-acting factors that cause this phenomenon. Can these cis- or trans-acting elements be recapitulated in other constitutively expressed messages in S2 cells and in mammalian cells?

References:
1. Shi Y., Giammartino D. C. D., Taylor D., Sarkeshik A., Rice W. J., Ill J. R. Y., Frank J., Manley J. L. (2009). Molecular architecture of the human pre-mRNA 3’ processing complex. Molecular Cell. 33: 365-376.
2. Nagaike T., Logan C., Hotta I., Rozenblatt-Rosen O., Meyerson M., Manley J. (2011). Transcriptional activators enhance polyadenylation of mRNA precursors. Molecular Cell. 41: 409-418.

Keywords: polyadenylation, polyA tail length assay