Specific trans-acting proteins interact with auxiliary RNA polyadenylation elements in the COX-2 3'-UTR

被引:68
作者
Hall-Pogar, Tyra [1 ]
Liang, Songchun [1 ]
Hague, Lisa K. [1 ]
Lutz, Carol S. [1 ]
机构
[1] Univ Med & Dent New Jersey, Grad Sch Biomed Sci, New Jersey Med Sch, Dept Biochem & Mol Biol, Newark, NJ 07103 USA
关键词
3'-untranslated region (UTR); polyadenylation; cyclooxygenase-2 (COX-2);
D O I
10.1261/rna.577707
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Two cyclooxygenase (COX) enzymes, COX-1 and COX-2, are present in human cells. While COX-1 is constitutively expressed, COX-2 is inducible and up-regulated in response to many signals. Since increased transcriptional activity accounts for only part of COX-2 up-regulation, we chose to explore other RNA processing mechanisms in the regulation of this gene. Previously, we showed that COX-2 is regulated by alternative polyadenylation, and that the COX-2 proximal polyadenylation signal contains auxiliary upstream sequence elements ( USEs) that are very important in efficient polyadenylation. To explore trans-acting protein factors interacting with these cis-acting RNA elements, we performed pull-down assays with HeLa nuclear extract and biotinylated RNA oligonucleotides representing COX-2 USEs. We identified PSF, p54(nrb), PTB, and U1A as proteins specifically bound to the COX-2 USEs. We further explored their participation in polyadenylation using MS2 phage coat protein-MS2 RNA binding site tethering assays, and found that tethering any of these four proteins to the COX-2 USE mutant RNA can compensate for these cis-acting elements. Finally, we suggest that these proteins (p54 nrb, PTB, PSF, and U1A) may interact as a complex since immunoprecipitations of the transfected MS2 fusion proteins coprecipitate the other proteins.
引用
收藏
页码:1103 / 1115
页数:13
相关论文
共 66 条
[1]   STRUCTURE OF THE HUMAN CYCLO-OXYGENASE-2 GENE [J].
APPLEBY, SB ;
RISTIMAKI, A ;
NEILSON, K ;
NARKO, K ;
HLA, T .
BIOCHEMICAL JOURNAL, 1994, 302 :723-727
[2]   COX-2 as a multifunctional neuronal modulator [J].
Bazan, NG .
NATURE MEDICINE, 2001, 7 (04) :414-415
[3]   PLATELET-ACTIVATING-FACTOR AND RETINOIC ACID SYNERGISTICALLY ACTIVATE THE INDUCIBLE PROSTAGLANDIN SYNTHASE GENE [J].
BAZAN, NG ;
FLETCHER, BS ;
HERSCHMAN, HR ;
MUKHERJEE, PK .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (12) :5252-5256
[4]   Rules of engagement: co-transcriptional recruitment of pre-mRNA processing factors [J].
Bentley, DL .
CURRENT OPINION IN CELL BIOLOGY, 2005, 17 (03) :251-256
[5]  
Bishop-Bailey David, 2002, Journal of Environmental Pathology Toxicology and Oncology, V21, P93
[6]   Cloning of guinea pig cyclooxygenase-2 and 15-hydroxyprostaglandin dehydrogenase complementary deoxyribonucleic acids: Steroid-modulated gene expression correlates to prostaglandin F-2 alpha secretion in cultured endometrial cells [J].
Bracken, KE ;
Elger, W ;
Jantke, I ;
Nanninga, A ;
Gellersen, B .
ENDOCRINOLOGY, 1997, 138 (01) :237-247
[7]   Recruitment of a basal polyadenylation factor by the upstream sequence element of the human lamin B2 polyadenylation signal [J].
Brackenridge, S ;
Proudfoot, NJ .
MOLECULAR AND CELLULAR BIOLOGY, 2000, 20 (08) :2660-2669
[8]   EFFICIENCY OF UTILIZATION OF THE SIMIAN VIRUS-40 LATE POLYADENYLATION SITE - EFFECTS OF UPSTREAM SEQUENCES [J].
CARSWELL, S ;
ALWINE, JC .
MOLECULAR AND CELLULAR BIOLOGY, 1989, 9 (10) :4248-4258
[9]   Polypyrimidine tract binding protein modulates efficiency of polyadenylation [J].
Castelo-Branco, P ;
Furger, A ;
Wollerton, M ;
Smith, C ;
Moreira, A ;
Proudfoot, N .
MOLECULAR AND CELLULAR BIOLOGY, 2004, 24 (10) :4174-4183
[10]   The 3′-untranslated region of murine cyclooxygenase-2 contains multiple regulatory elements that alter message stability and translational efficiency [J].
Cok, SJ ;
Morrison, AR .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (25) :23179-23185