The 3′-untranslated region of p21WAF1 mRNA is a composite cis-acting sequence bound by RNA-binding proteins from breast cancer cells, including HuR and poly(C)-binding protein

被引:98
作者
Giles, KM
Daly, JM
Beveridge, DJ
Thomson, AM
Voon, DC
Furneaux, HM
Jazayeri, JA
Leedman, PJ
机构
[1] Univ Western Australia, Lab Canc Med, Perth, WA 6001, Australia
[2] Univ Western Australia, Dept Med, Western Australian Inst Med Res, Perth, WA 6001, Australia
[3] Univ Western Australia, Med Res Ctr, Perth, WA 6001, Australia
[4] Royal Perth Hosp, Dept Endocrinol & Diabet, Perth, WA 6001, Australia
[5] Univ Connecticut, Ctr Hlth, Dept Physiol, Vasc Biol Program, Farmington, CT 06030 USA
关键词
D O I
10.1074/jbc.M208439200
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
Despite promoting growth in many cell types, epidermal growth factor (EGF) induces growth inhibition in a variety of cancer cells that overexpress its receptor. The cyclin-dependent kinase inhibitor p21(WAF1) is a central component of this pathway. We found in human MDA-468 breast cancer cells that EGF up-regulates p21(WAF1) mRNA and protein, through a combination of increased mRNA stability and transcription. The decay rate of a hybrid luciferase reporter full-length p21(WAF1) 3'-untranslated region (UTR) mRNA was significantly faster than that of a control mRNA. Transfections with a variety of p21(WAF1) 3'-UTR constructs identified multiple cis-acting elements capable of reducing basal reporter activity. Short wavelength ultraviolet light induced reporter activity in constructs containing the 5' region of the p21(WAF1) 3'-UTR, whereas EGF induced reporter activity in constructs containing sequences 3' of the UVC-responsive region. These cis-elements bound multiple proteins from MDA-468 cells, including HuR and poly(C)-binding protein 1 (CP1). Immunoprecipitation studies confirmed that HuR and CP1 associate with p21(WAF1) mRNA in MDA-468 cells. Over- and underexpression of HuR in MDA-468 cells did not affect EGF-induced p21(WAF1) protein expression or growth inhibition. However, binding of HuR to its target 3'-UTR cis-element was regulated by UVC but not by EGF, suggesting that these stimuli modulate the stability of p21(WAF1) mRNA via different mechanisms. We conclude that EGF-induced p21(WAF1) protein expression is mediated largely by stabilization of p21(WAF1) mRNA elicited via multiple 3'-UTR cis-elements. Although HuR binds at least one of these elements, it does not appear to be a major modulator of p21(WAF1) expression or growth inhibition in this system. CP1 is a novel p21(WAF1) mRNA-binding protein that may function cooperatively with other mRNA-binding proteins to regulate p21(WAF1) mRNA stability.
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页码:2937 / 2946
页数:10
相关论文
共 61 条
  • [51] SHEIKH MS, 1995, ONCOGENE, V11, P1899
  • [52] Shiohara M, 1996, J CELL PHYSIOL, V166, P568, DOI 10.1002/(SICI)1097-4652(199603)166:3<568::AID-JCP11>3.0.CO
  • [53] 2-3
  • [54] The inhibitory activity of the AU-rich RNA element in the human papillomavirus type 1 late 3′ untranslated region correlates with its affinity for the elav-like HuR protein
    Sokolowski, M
    Furneaux, E
    Schwartz, S
    [J]. JOURNAL OF VIROLOGY, 1999, 73 (02) : 1080 - 1091
  • [55] Optimized RNA gel-shift and UV cross-linking assays for characterization of cytoplasmic RNA-protein interactions
    Thomson, AM
    Rogers, JT
    Walker, CE
    Staton, JM
    Leedman, PJ
    [J]. BIOTECHNIQUES, 1999, 27 (05) : 1032 - +
  • [56] HuR regulates p21 mRNA stabilization by UV light
    Wang, WG
    Furneaux, H
    Cheng, HM
    Caldwell, MC
    Hutter, D
    Liu, YS
    Holbrook, N
    Gorospe, M
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 2000, 20 (03) : 760 - 769
  • [57] WANG XM, 1995, MOL CELL BIOL, V15, P1769
  • [58] Xie W, 1997, ANTICANCER RES, V17, P2627
  • [59] Differential posttranscriptional regulation of androgen receptor gene expression by androgen in prostate and breast cancer cells
    Yeap, BB
    Krueger, RG
    Leedman, PJ
    [J]. ENDOCRINOLOGY, 1999, 140 (07) : 3282 - 3291
  • [60] Novel binding of HuR and poly(C)-binding protein to a conserved UC-rich motif within the 3′-untranslated region of the androgen receptor messenger RNA
    Yeap, BB
    Voon, DC
    Vivian, JP
    McCulloch, RK
    Thomson, AM
    Giles, KM
    Czyzyk-Krzeska, MF
    Furneaux, H
    Wilce, MCJ
    Wilce, JA
    Leedman, PJ
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (30) : 27183 - 27192