Structural insights into the transcriptional and translational roles of Ebp1.
The ErbB3-binding protein 1 (Ebp1) is an important regulator of transcription, affecting eukaryotic cell growth, proliferation, differentiation and survival. Ebp1 can also affect translation and cooperates with the polypyrimidine tract-binding protein (PTB) to stimulate the activity of the internal ribosome entry site (IRES) of foot-and-mouth disease virus (FMDV). We report here the crystal structure of murine Ebp1 (p48 isoform), providing the first glimpse of the architecture of this versatile regulator. The structure reveals a core domain that is homologous to methionine aminopeptidases, coupled to a C-terminal extension that contains important motifs for binding proteins and RNA. It sheds new light on the conformational differences between the p42 and p48 isoforms of Ebp1, the disposition of the key protein-interacting motif ((354)LKALL(358)) and the RNA-binding activity of Ebp1. We show that the primary RNA-binding site is formed by a Lys-rich motif in the C terminus and mediates the interaction with the FMDV IRES. We also demonstrate a specific functional requirement for Ebp1 in FMDV IRES-directed translation that is independent of a direct interaction with PTB.
Monie
Tom P
TP
Division of Cell and Molecular Biology, Imperial College, South Kensington Campus, London, UK.
Perrin
Andrew J
AJ
Birtley
James R
JR
Sweeney
Trevor R
TR
Karakasiliotis
Ioannis
I
Chaudhry
Yasmin
Y
Roberts
Lisa O
LO
Matthews
Stephen
S
Goodfellow
Ian G
IG
Curry
Stephen
S
D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000502:physiology; D000626:Aminopeptidases / Q000737:chemistry; D001665:Binding Sites; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics; D008239:Lysine / Q000737:chemistry; D063208:Methionyl Aminopeptidases; D008958:Models, Molecular; D014176:Protein Biosynthesis; D011487:Protein Conformation; D020033:Protein Isoforms / Q000737:chemistry / Q000502:physiology; D034741:RNA, Small Interfering / Q000235:genetics; D012367:RNA, Viral / Q000737:chemistry; D015533:Transcriptional Activation
D048868:Adaptor Proteins, Signal Transducing; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D012367:RNA, Viral; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases; D008239:Lysine
Nucleic Acids Res. 2005;33(18):6024-33
16254079
Trends Biochem Sci. 2005 Feb;30(2):66-9
15691650
J Mol Biol. 2005 Jul 1;350(1):145-55
15919091
Mol Cell. 2002 Feb;9(2):329-39
11864606
Biochem Soc Trans. 2005 Dec;33(Pt 6):1231-41
16246087
EMBO J. 2000 Jun 15;19(12):3132-41
10856256
EMBO J. 2006 Oct 18;25(20):4909-20
17024182
Chem Rev. 2002 Dec;102(12):4581-608
12475202
Nucleic Acids Res. 2003 Apr 15;31(8):2168-77
12682367
Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2473-7
8146141
EMBO J. 2006 May 17;25(10):2083-95
16642037
Cytogenet Cell Genet. 1997;78(1):31-5
9345902
Oncogene. 2004 Apr 19;23(18):3200-7
15094769
J Cell Physiol. 2000 Jun;183(3):321-9
10797306
Proc Natl Acad Sci U S A. 2006 Jul 18;103(29):10917-22
16832058
Exp Cell Res. 1995 Oct;220(2):434-45
7556453
Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6350-5
10829067
Biochem Biophys Res Commun. 2006 Jun 9;344(3):859-68
16631606
Acta Crystallogr D Biol Crystallogr. 2004 Mar;60(Pt 3):432-8
14993666
Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21
9757107
Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9
2025413
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2256-68
15572779
J Virol. 1988 Aug;62(8):2636-43
2839690
Methods Enzymol. 2003;374:229-44
14696376
Proc Natl Acad Sci U S A. 2005 Jul 12;102(28):9890-5
15994225
Genes Dev. 2000 Aug 15;14(16):2028-45
10950867
Nat Struct Mol Biol. 2006 Sep;13(9):839-48
16936729
EMBO J. 2003 Dec 1;22(23):6356-64
14633994
Br J Cancer. 2000 Feb;82(3):683-90
10682683
Oncogene. 2002 Aug 15;21(36):5609-18
12165860
J Cell Physiol. 2001 May;187(2):209-17
11268000
Mol Cell. 2003 Mar;11(3):757-71
12667457
Nature. 1998 Sep 10;395(6698):137-43
9744270
EMBO Rep. 2005 Feb;6(2):140-6
15654320
RNA. 1998 May;4(5):520-9
9582094
J Biol Chem. 2004 Jun 18;279(25):26126-33
15073182
J Mol Biol. 1998 Nov 20;284(1):101-24
9811545
Mol Cell Endocrinol. 2001 Apr 25;175(1-2):185-91
11325528
Yeast. 1994 Jul;10(7):883-94
7985416
J Virol. 1994 Feb;68(2):697-703
8289373
Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7029-36
11416183
Structure. 2004 Sep;12(9):1631-43
15341728
Biochemistry. 1997 Sep 30;36(39):11881-90
9305981
RNA. 2001 Jan;7(1):114-22
11214173
Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3
15299374
Nature. 1988 Jul 28;334(6180):320-5
2839775
Oncogene. 2004 May 27;23(25):4454-65
15064750
PLoS Biol. 2004 Sep;2(9):E274
15328534
Exp Cell Res. 2006 Jun 10;312(10):1703-12
16515785
Genes Dev. 2001 Jul 1;15(13):1593-612
11445534
Bioorg Med Chem Lett. 2006 May 15;16(10):2580-3
16540317
Nucleic Acids Res. 2003 Jul 1;31(13):3381-5
12824332
Science. 1998 Nov 13;282(5392):1324-7
9812898
Genes Dev. 1999 Feb 15;13(4):437-48
10049359
Biochem J. 2006 May 15;396(1):99-107
16396631