Histone mRNA levels are tightly coordinated with DNA replication to ensure proper replication of chromatin. The replication-dependent histone genes encode the only non-polyadenylated mRNAs is eukaryotic cells. These mRNAs end in a conserved stemloop, which interacts with the stemloop binding protein, SLBP. The stem-loop/SLBP complex functions in all steps of histone mRNA metabolism in place of the polyA tail. Most of the regulation of histone mRNA levels is posttranscriptional and is mediated by the stemloop/SLBP complex. Formation of the 3'end of histone mRNA is cell-cycle regulated, primarily mediated by controlling the levels of SLBP. We will determine the additional components that recruit the cleavage factor to the histone pre-mRNA pre-processing complex. There are likely factors that regulate processing that influence this recruitment and we will characterize both positive and negative factors, involved in this critical regulatory step. The second regulatory step that coordinates histone mRNA levels with DNA replication is regulation of histone mRNA half-life. Histone mRNA degradation is initiated by oligouridylation of the 3'end. We will identify the terminal uridyl transferase (TUTase) involved in the oligouridylation, and the mechanism by which the TUTase is recruited to activate histone mRNA degradation. These studies will allow us to identify novel factors and modifications involved in coupling DNA replication with histone mRNA degradation.

Public Health Relevance

Each S-phase the cell must properly replicate its genome, and package the newly replicated DNA into chromatin. The levels of histone mRNA are tightly controlled by regulating both 3'end formation and mRNA degradation. We will determine the biochemical mechanisms that couple histone mRNA metabolism with DNA replication.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM029832-38
Application #
8478120
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Bender, Michael T
Project Start
1982-07-01
Project End
2015-05-31
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
38
Fiscal Year
2013
Total Cost
$376,826
Indirect Cost
$121,485
Name
University of North Carolina Chapel Hill
Department
Biochemistry
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Lackey, Patrick E; Welch, Joshua D; Marzluff, William F (2016) TUT7 catalyzes the uridylation of the 3' end for rapid degradation of histone mRNA. RNA 22:1673-1688
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Borchardt, Erin K; Vandoros, Leonidas A; Huang, Michael et al. (2015) Controlling mRNA stability and translation with the CRISPR endoribonuclease Csy4. RNA 21:1921-30
Welch, Joshua D; Slevin, Michael K; Tatomer, Deirdre C et al. (2015) EnD-Seq and AppEnD: sequencing 3' ends to identify nontemplated tails and degradation intermediates. RNA 21:1375-89
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Zhang, Jun; Tan, Dazhi; DeRose, Eugene F et al. (2014) Molecular mechanisms for the regulation of histone mRNA stem-loop-binding protein by phosphorylation. Proc Natl Acad Sci U S A 111:E2937-46
Djakbarova, Umidahan; Marzluff, William F; Koseoglu, M Murat (2014) Translation regulation and proteasome mediated degradation cooperate to keep stem-loop binding protein low in G1-phase. J Cell Biochem 115:523-30
Lyons, Shawn M; Ricciardi, Adele S; Guo, Andrew Y et al. (2014) The C-terminal extension of Lsm4 interacts directly with the 3' end of the histone mRNP and is required for efficient histone mRNA degradation. RNA 20:88-102
Slevin, Michael K; Meaux, Stacie; Welch, Joshua D et al. (2014) Deep sequencing shows multiple oligouridylations are required for 3' to 5' degradation of histone mRNAs on polyribosomes. Mol Cell 53:1020-30

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