We have a long-standing interest in how eukaryotic mRNA 3' ends are formed. Regulation of gene expression can be accomplished by regulating polyadenylation. We have combined this interest in polyadenylation with an interest in human collagen types I and II. Collagen plays a key role in normal cartilage development and skeletal morphogenesis. We have recently discovered and published that human COL1A2 polyadenylation is regulated by cis-acting elements present in the 3' untranslated region (3' UTR) of the COL1A2 mRNA, and that these elements influence the 3' end processing efficiency. Human COL1A1 and COL2A1 we have now discovered also contain these cis-acting elements. Based upon these findings, this project proposes to investigate the mechanism of regulation of polyadenylation of all these type I and II collagen pre-mRNAs. These investigations will expand our goal of better understanding the expression of collagens, and how mutations and abnormal gene expression regulation can initiate a disease state.
Our specific aims for this project are: 1) To identify all the cis-acting elements involved in 3' polyadenylation site usage in the human collagen genes COL1A1, COL1A2, and COL2A1 and 2) To determine what trans-acting factor(s) bind to these cis-acting elements. First, we will identify cis-acting elements by deletion and site-directed mutagenesis, and examine their ability to affect polyadenylation efficiency both in vitro and in vivo. Second, protein factors that bind to these cis-acting signals will be explored and characterized. Determination of these protein factors will provide us with clues as to the functional significance of cis-acting auxiliary elements. Gaining an understanding of the RNA signals and protein factors involved in regulation of 3' end formation of these human collagen genes will help us to comprehend collagen gene expression regulation, which will in turn lead to novel insights into skeletal defects and other collagen-based diseases. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Small Research Grants (R03)
Project #
5R03AR052038-03
Application #
7169265
Study Section
Special Emphasis Panel (ZAR1-YZW-E (O2))
Program Officer
Tyree, Bernadette
Project Start
2005-01-15
Project End
2008-11-30
Budget Start
2006-12-01
Budget End
2008-11-30
Support Year
3
Fiscal Year
2007
Total Cost
$73,721
Indirect Cost
Name
University of Medicine & Dentistry of NJ
Department
Biochemistry
Type
Schools of Medicine
DUNS #
623946217
City
Newark
State
NJ
Country
United States
Zip Code
07107
Lutz, Carol S (2008) Alternative polyadenylation: a twist on mRNA 3'end formation. ACS Chem Biol 3:609-17
Hague, Lisa K; Hall-Pogar, Tyra; Lutz, Carol S (2008) In vivo methods to assess polyadenylation efficiency. Methods Mol Biol 419:171-85
Liang, Songchun; Lutz, Carol S (2006) p54nrb is a component of the snRNP-free U1A (SF-A) complex that promotes pre-mRNA cleavage during polyadenylation. RNA 12:111-21
Nemeth, Zoltan H; Lutz, Carol S; Csoka, Balazs et al. (2005) Adenosine augments IL-10 production by macrophages through an A2B receptor-mediated posttranscriptional mechanism. J Immunol 175:8260-70
Hu, Jun; Lutz, Carol S; Wilusz, Jeffrey et al. (2005) Bioinformatic identification of candidate cis-regulatory elements involved in human mRNA polyadenylation. RNA 11:1485-93