Hairpin structure within the 3'UTR of DNA polymerase beta mRNA acts as a post-transcriptional regulatory element and interacts with Hax-1.
Aberrant expression of DNA polymerase beta, a key enzyme involved in base excision repair, leads to genetic instability and carcinogenesis. Pol beta expression has been previously shown to be regulated at the level of transcription, but there is also evidence of post-transcriptional regulation, since rat transcripts undergo alternative polyadenylation, and the resulting 3'UTR contain at least one regulatory element. Data presented here indicate that RNA of the short 3'UTR folds to form a strong secondary structure (hairpin). Its regulatory role was established utilizing a luciferase-based reporter system. Further studies led to the identification of a protein factor, which binds to this element-the anti-apoptotic, cytoskeleton-related protein Hax-1. The results of in vitro binding analysis indicate that the formation of the RNA-protein complex is significantly impaired by disruption of the hairpin motif. We demonstrate that Hax-1 binds to Pol beta mRNA exclusively in the form of a dimer. Biochemical analysis revealed the presence of Hax-1 in mitochondria, but also in the nuclear matrix, which, along with its transcript-binding properties, suggests that Hax-1 plays a role in post-transcriptional regulation of expression of Pol beta.
Sarnowska
Elzbieta
E
Cancer Center Institute, Roentgena 5, 02-781 Warsaw, Poland.
Grzybowska
Ewa A
EA
Sobczak
Krzysztof
K
Konopinski
Ryszard
R
Wilczynska
Anna
A
Szwarc
Maria
M
Sarnowski
Tomasz J
TJ
Krzyzosiak
Wlodzimierz J
WJ
Siedlecki
Janusz A
JA
D020413:3' Untranslated Regions / Q000737:chemistry*; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism*; D045744:Cell Line, Tumor; D019951:DNA Polymerase beta / Q000235:genetics*; D019281:Dimerization; D019143:Evolution, Molecular; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D047908:Intracellular Signaling Peptides and Proteins; D008928:Mitochondria / Q000737:chemistry; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015530:Nuclear Matrix / Q000737:chemistry; D009690:Nucleic Acid Conformation; D051381:Rats; D038621:Regulatory Sequences, Ribonucleic Acid; D014466:Ultraviolet Rays
D020413:3' Untranslated Regions; D002352:Carrier Proteins; C510653:Hax1 protein, rat; D047908:Intracellular Signaling Peptides and Proteins; D038621:Regulatory Sequences, Ribonucleic Acid; D019951:DNA Polymerase beta
EMBO J. 2000 Mar 15;19(6):1397-404
10716939
Proc Natl Acad Sci U S A. 1998 Oct 13;95(21):12586-90
9770529
J Cell Sci. 2000 Jul;113 ( Pt 13):2433-43
10852822
J Virol. 2000 Nov;74(21):10104-11
11024139
Oncogene. 2000 Sep 28;19(41):4729-35
11032023
J Biol Chem. 2001 Apr 27;276(17):13593-9
11139568
Prog Nucleic Acid Res Mol Biol. 2001;68:57-74
11554313
Cytokine. 2001 Aug 7;15(3):122-37
11554782
Oncogene. 2001 Sep 27;20(43):6181-7
11593426
J Virol. 2002 Jan;76(2):802-16
11752170
Genome Res. 2002 Apr;12(4):656-64
11932250
Cancer Res. 2002 Jun 15;62(12):3511-4
12067997
Nucleic Acids Res. 2001 May 1;29(9):e45
11328886
Nucleic Acids Res. 2002 Dec 1;30(23):5017-28
12466525
Biol Chem. 2003 Jan;384(1):19-23
12674496
Nucleic Acids Res. 2003 Jul 1;31(13):3406-15
12824337
J Biol Chem. 2004 Jul 30;279(31):32761-70
15159385
Nucleic Acids Res. 1978 Nov;5(11):4263-72
724514
Gene. 1987;57(2-3):267-72
3319781
Science. 1988 May 13;240(4854):924-8
2452485
Exp Cell Res. 1989 Mar;181(1):169-80
2917600
Biochim Biophys Acta. 1989 Jul 7;1008(2):203-7
2736248
J Biol Chem. 1991 May 25;266(15):9893-9
2033075
Yeast. 1991 Jul;7(5):475-7
1897313
Curr Genet. 1992 Jan;21(1):83-4
1735128
Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5428-32
8516284
RNA. 1996 Apr;2(4):367-79
8634917
Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8496-501
8710898
Yeast. 1997 Sep 15;13(11):1065-75
9290211
Cancer Res. 1999 Apr 1;59(7):1544-51
10197627
Carcinogenesis. 1999 Jun;20(6):1049-54
10357787
J Biol Chem. 2004 Nov 19;279(47):49406-13
15339924
J Biol Chem. 2004 Nov 26;279(48):50295-301
15371414
Brief Funct Genomic Proteomic. 2004 Nov;3(3):240-56
15642187
RNA. 2005 Mar;11(3):294-307
15701732
Biochem J. 2005 Apr 15;387(Pt 2):419-28
15537387
Biochem J. 2005 Dec 15;392(Pt 3):475-83
16042622
Proc Natl Acad Sci U S A. 2006 Sep 26;103(39):14501-6
16971486
Biochemistry. 2006 Dec 12;45(49):14632-41
17144656
J Biol Chem. 1996 Aug 2;271(31):18508-13
8702497
Gene. 1996 Oct 17;176(1-2):191-5
8918252
J Immunol. 1997 Mar 15;158(6):2736-44
9058808
J Cell Biol. 1997 Apr 21;137(2):263-74
9128241
Nucleic Acids Res. 1997 Aug 15;25(16):3362-70
9241253
Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):4017-22
10760273