This research is aimed at understanding the regulation of gene expression in animal cells with specific emphasis on transcription and processing the primary nuclear transcripts (hnRNA) into messenger RNA (mRNA) HnRNA contains a large number of oligo U sequences that are conserved in a significant fraction (15-20%) of the polyadenylated mRNA's of the cytoplasm. The structure of these mRNA's, their translatability and sequence relationships to mRNA's lacking oligo U's are being studied. Nuclear polyadenylated RNA also contain branched structures formed by a phosphodiester bond at the 3 feet hydroxyl group of a nucleoside that is already linked via the normal 3 feet 5 feet phosphodiester bond to an adjacent nucleotide. The absence of branched structures from polyadenylated mRNA leads us to suspect such structures are involved in a nuclear RNA processing reaction. Structural analysis and localization and distribution of branches in RNA sub-populations are proposed. Branches will be sought in adenovirus primary transcripts to develop a well-defined system for studying the possible role of branches in nuclear RNA processing.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032585-03
Application #
3281561
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1983-09-01
Project End
1988-08-31
Budget Start
1985-09-01
Budget End
1986-08-31
Support Year
3
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Pittsburgh
Department
Type
Schools of Arts and Sciences
DUNS #
053785812
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Reilly, J D; Melhem, R F; Lutz, C M et al. (1990) Transcription vectors that facilitate the identification and mapping of RNA splice sites in genomic DNA. DNA Cell Biol 9:535-42
Reilly, J D; Wallace, J C; Edmonds, M (1987) The quantitation and distribution of splicing intermediates in HeLa cells and adenovirus RNAs. Nucleic Acids Res 15:7103-24
Kierzek, R; Kopp, D W; Edmonds, M et al. (1986) Chemical synthesis of branched RNA. Nucleic Acids Res 14:4751-64
Wood, W M; Wallace, J C; Edmonds, M (1985) Sequence content of oligo(uridylic acid)-containing messenger ribonucleic acid from HeLa cells. Biochemistry 24:3686-93
Kulkosky, J W; Wood, W M; Edmonds, M (1985) Location of oligo(uridylic acid) sequences within messenger ribonucleic acid molecules of HeLa cells. Biochemistry 24:3678-86