Structural insights into the interaction of the evolutionarily conserved ZPR1 domain tandem with eukaryotic EF1A, receptors, and SMN complexes.
Eukaryotic genomes encode a zinc finger protein (ZPR1) with tandem ZPR1 domains. In response to growth stimuli, ZPR1 assembles into complexes with eukaryotic translation elongation factor 1A (eEF1A) and the survival motor neurons protein. To gain insight into the structural mechanisms underlying the essential function of ZPR1 in diverse organisms, we determined the crystal structure of a ZPR1 domain tandem and characterized the interaction with eEF1A. The ZPR1 domain consists of an elongation initiation factor 2-like zinc finger and a double-stranded beta helix with a helical hairpin insertion. ZPR1 binds preferentially to GDP-bound eEF1A but does not directly influence the kinetics of nucleotide exchange or GTP hydrolysis. However, ZPR1 efficiently displaces the exchange factor eEF1Balpha from preformed nucleotide-free complexes, suggesting that it may function as a negative regulator of eEF1A activation. Structure-based mutational and complementation analyses reveal a conserved binding epitope for eEF1A that is required for normal cell growth, proliferation, and cell cycle progression. Structural differences between the ZPR1 domains contribute to the observed functional divergence and provide evidence for distinct modalities of interaction with eEF1A and survival motor neuron complexes.
Mishra
Ashwini K
AK
Program in Molecular Medicine and Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01655, USA.
Gangwani
Laxman
L
Davis
Roger J
RJ
Lambright
David G
DG
D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017124:Conserved Sequence; D004252:DNA Mutational Analysis; D019143:Evolution, Molecular; D020558:GTP Phosphohydrolases / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D008958:Models, Molecular; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics
D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D011994:Recombinant Proteins; C490987:Zfp259 protein, mouse; D006153:Guanosine Diphosphate; D020558:GTP Phosphohydrolases
Acta Crystallogr D Biol Crystallogr. 1999 Apr;55(Pt 4):849-61
10089316
Hum Mol Genet. 2000 Feb 12;9(3):341-6
10655542
Nat Cell Biol. 2001 Apr;3(4):376-83
11283611
Mol Cell Biol. 2002 Oct;22(20):6993-7003
12242280
Nat Struct Biol. 2002 Dec;9(12):900-5
12402030
Biochemistry. 2003 Aug 19;42(32):9515-24
12911293
Hum Genet. 2003 Dec;114(1):11-21
14520560
Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3
15299374
Acta Crystallogr D Biol Crystallogr. 1996 Jan 1;52(Pt 1):30-42
15299723
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2256-68
15572779
Hum Mol Genet. 2005 Mar 15;14(6):845-57
15703193
Mol Cell Biol. 2005 Apr;25(7):2744-56
15767679
J Neuropathol Exp Neurol. 2005 Apr;64(4):295-303
15835265
Nat Struct Mol Biol. 2005 Sep;12(9):772-8
16116436
Neuron. 2005 Dec 22;48(6):885-96
16364894
Nucleic Acids Res. 2006 Jan 1;34(Database issue):D257-60
16381859
Biochim Biophys Acta. 2006 Jan-Feb;1759(1-2):13-31
16624425
Proc Natl Acad Sci U S A. 2006 May 9;103(19):7471-5
16648254
J Biol Chem. 2006 Dec 29;281(52):40330-40
17068332
Mol Cell Biol. 2007 Mar;27(5):1974-89
17178834
Methods Enzymol. 1991;194:302-18
2005795
Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9
2025413
Methods Enzymol. 1997;276:307-26
27754618
Methods Enzymol. 1997;276:472-494
27799110
J Biol Chem. 1988 Feb 5;263(4):1773-8
3338993
Cell. 1995 Jan 13;80(1):155-65
7813012
Eur J Biochem. 1993 Aug 1;215(3):549-54
8354261
Science. 1996 Jun 21;272(5269):1797-802
8650580
Genomics. 1996 Sep 1;36(2):359-61
8812466
Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4463-8
9539760
Mol Biol Cell. 1998 Oct;9(10):2963-71
9763455
J Cell Biol. 1998 Dec 14;143(6):1471-84
9852145