Ultraviolet A-induced modulation of Bcl-XL by p38 MAPK in human Keratinocytes -: Post-transcriptional regulation through the 3′-untranslated region

被引:43
作者
Bachelor, MA [1 ]
Bowden, GT [1 ]
机构
[1] Univ Arizona, Arizona Canc Ctr, Dept Cell Biol & Anat, Tucson, AZ 85724 USA
关键词
D O I
10.1074/jbc.M406626200
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
We examined the effect of inhibiting p38 MAPK on UVA-irradiated HaCaT cells, a spontaneously immortalized human keratinocyte cell line. Recent work from our laboratory has shown that UVA ( 250 kJ/m(2)) induces a rapid phosphorylation of p38 MAPK in the HaCaT cell line. Inhibition of p38 MAPK activity through the use of a specific inhibitor, SB202190, in combination with UVA treatment induced a rapid cleavage of caspase-9, caspase-8, and caspase-3, whereas UVA irradiation alone had no effect. Similarly, cleavage of the caspase substrate poly( ADPribose) polymerase was observed in UVA-irradiated HaCaT cells treated with SB202190 or in cells expressing a dominant-negative p38 MAPK. No effect of p38 MAPK inhibition upon caspase cleavage was observed in mock-irradiated HaCaT cells. In addition, increases in apoptosis were observed in UVA-irradiated cells treated with SB202190 by morphological analysis with no significant apoptosis occurring from UVA irradiation alone. Similar results were obtained by using normal human epidermal keratinocytes. UVA induced expression of the anti-apoptotic Bcl-2 family member, Bcl-X-L, with abrogation of expression by using the p38 MAPK inhibitor SB202190. Overexpression of Bcl-X-L prevented poly(ADP-ribose) polymerase cleavage induced by the combination of UVA and p38 MAPK inhibition. UVA enhanced the stability of Bcl-X-L mRNA through increases in p38 MAPK activity. We determined that increases in UVA-induced expression of Bcl-X-L occur through a posttranscriptional mechanism mediated by the 3'-untranslated region (UTR). We used Bcl-X-L 3'-UTR luciferase constructs to determine the mechanism by which UVA increased Bcl-X-L mRNA stability. Additionally, RNA binding studies indicate that UVA increases the binding of RNA-binding proteins to Bcl-X-L 3'-UTR mRNA, which can be decreased by using SB202190. In conclusion, p38 MAPK and Bcl-X-L expression play critical roles in the survival of UVA-irradiated HaCaT cells.
引用
收藏
页码:42658 / 42668
页数:11
相关论文
共 53 条
[1]   The role of p38 in UVA-induced cyclooxygenase-2 expression in the human keratinocyte cell line, HaCaT [J].
Bachelor, MA ;
Silvers, AL ;
Bowden, GT .
ONCOGENE, 2002, 21 (46) :7092-7099
[2]   UVA exposure of human skin reconstructed in vitro induces apoptosis of dermal fibroblasts:: subsequent connective tissue repair and implications in photoaging [J].
Bernerd, F ;
Asselineau, D .
CELL DEATH AND DIFFERENTIATION, 1998, 5 (09) :792-802
[3]   AUTOANTIGENS TARGETED IN SYSTEMIC LUPUS-ERYTHEMATOSUS ARE CLUSTERED IN 2 POPULATIONS OF SURFACE-STRUCTURES ON APOPTOTIC KERATINOCYTES [J].
CASCIOLAROSEN, LA ;
ANHALT, G ;
ROSEN, A .
JOURNAL OF EXPERIMENTAL MEDICINE, 1994, 179 (04) :1317-1330
[4]   ABERRANT BCL-2 PROTEIN EXPRESSION PROVIDES A POSSIBLE MECHANISM OF NEOPLASTIC CELL-GROWTH IN CUTANEOUS BASAL-CELL CARCINOMA [J].
CERRONI, L ;
KERL, H .
JOURNAL OF CUTANEOUS PATHOLOGY, 1994, 21 (05) :398-403
[5]   SELECTIVE DEGRADATION OF EARLY-RESPONSE-GENE MESSENGER-RNAS - FUNCTIONAL ANALYSES OF SEQUENCE FEATURES OF THE AU-RICH ELEMENTS [J].
CHEN, CYA ;
SHYU, AB .
MOLECULAR AND CELLULAR BIOLOGY, 1994, 14 (12) :8471-8482
[6]   AU-RICH ELEMENTS - CHARACTERIZATION AND IMPORTANCE IN MESSENGER-RNA DEGRADATION [J].
CHEN, CYA ;
SHYU, AB .
TRENDS IN BIOCHEMICAL SCIENCES, 1995, 20 (11) :465-470
[7]   Activation of p38 MAP kinase and ERK are required for ultraviolet-B induced c-fos gene expression in human keratinocytes [J].
Chen, WX ;
Bowden, GT .
ONCOGENE, 1999, 18 (52) :7469-7476
[8]   UVB-mediated activation of p38 mitogen-activated protein kinase enhances resistance of normal human keratinocytes to apoptosis by stabilizing cytoplasmic p53 [J].
Chouinard, N ;
Valerie, K ;
Rouabhia, M ;
Huot, J .
BIOCHEMICAL JOURNAL, 2002, 365 (01) :133-145
[9]   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
[10]  
de Gruijl FR, 2000, METHOD ENZYMOL, V319, P359