Structural basis of E2-25K/UBB+1 interaction leading to proteasome inhibition and neurotoxicity.
E2-25K/Hip2 is an unusual ubiquitin-conjugating enzyme that interacts with the frameshift mutant of ubiquitin B (UBB(+1)) and has been identified as a crucial factor regulating amyloid-β neurotoxicity. To study the structural basis of the neurotoxicity mediated by the E2-25K-UBB(+1) interaction, we determined the three-dimensional structures of UBB(+1), E2-25K and the E2-25K/ubiquitin, and E2-25K/UBB(+1) complex. The structures revealed that ubiquitin or UBB(+1) is bound to E2-25K via the enzyme MGF motif and residues in α9 of the enzyme. Polyubiquitylation assays together with analyses of various E2-25K mutants showed that disrupting UBB(+1) binding markedly diminishes synthesis of neurotoxic UBB(+1)-anchored polyubiquitin. These results suggest that the interaction between E2-25K and UBB(+1) is critical for the synthesis and accumulation of UBB(+1)-anchored polyubiquitin, which results in proteasomal inhibition and neuronal cell death.
Ko
Sunggeon
S
Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749, Korea.
Kang
Gil Bu
GB
Song
Sung Min
SM
Lee
Jung-Gyu
JG
Shin
Dong Yeon
DY
Yun
Ji-Hye
JH
Sheng
Yi
Y
Cheong
Chaejoon
C
Jeon
Young Ho
YH
Jung
Yong-Keun
YK
Arrowsmith
Cheryl H
CH
Avvakumov
George V
GV
Dhe-Paganon
Sirano
S
Yoo
Yung Joon
YJ
Eom
Soo Hyun
SH
Lee
Weontae
W
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D045744:Cell Line, Tumor; D002470:Cell Survival; D018360:Crystallography, X-Ray; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D006860:Hydrogen Bonding; D009682:Magnetic Resonance Spectroscopy; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism*; D009154:Mutation; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D025801:Ubiquitin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D046912:Multiprotein Complexes; D011993:Recombinant Fusion Proteins; C479419:UBB protein, human; D025801:Ubiquitin; D049452:Green Fluorescent Proteins; C100879:UBE2K protein, human; D044763:Ubiquitin-Conjugating Enzymes; D046988:Proteasome Endopeptidase Complex
Proc Natl Acad Sci U S A. 2000 Aug 29;97(18):9902-6
10944193
Nat Struct Mol Biol. 2005 Aug;12(8):708-14
16007098
J Biol Chem. 1990 Dec 15;265(35):21835-42
2174887
J Mol Biol. 2006 May 5;358(3):713-24
16563434
J Alzheimers Dis. 2006;9(3 Suppl):319-25
16914870
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 May 1;65(Pt 5):440-4
19407372
PLoS Genet. 2009 Feb;5(2):e1000382
19214209
Mol Cell Neurosci. 2007 Jan;34(1):10-9
17092742
Structure. 2005 Apr;13(4):521-32
15837191
Proc Natl Acad Sci U S A. 2007 Sep 4;104(36):14336-41
17726107
J Cell Mol Med. 2008 Apr;12(2):363-73
18266959
J Biol Chem. 2005 Sep 9;280(36):31732-8
16014632
Proc Natl Acad Sci U S A. 2009 Jul 21;106(29):11907-12
19581590
J Biomol NMR. 1999 Mar;13(3):289-302
10212987
Annu Rev Neurosci. 2001;24:1121-59
11520930
J Cell Biol. 2002 Apr 29;157(3):417-27
11980917
Mol Cell. 2003 Sep;12(3):553-63
14527403
Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
15299926
J Biol Chem. 2004 Nov 5;279(45):47139-47
15328341
Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21
9757107
J Biol Chem. 2007 Dec 7;282(49):35787-95
17897937
Biochemistry. 1997 Aug 26;36(34):10526-37
9265633
Science. 1998 Jan 9;279(5348):242-7
9422699
Neuron. 2001 Jan;29(1):15-32
11182078
Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12366-71
9356455
Methods Enzymol. 1997;276:307-26
27754618
Nat Rev Mol Cell Biol. 2005 Aug;6(8):610-21
16064137
J Cell Biol. 2008 Aug 25;182(4):675-84
18710920
J Mol Graph. 1996 Feb;14(1):51-5, 29-32
8744573
Science. 2001 May 25;292(5521):1552-5
11375494
J Biomol NMR. 1995 Nov;6(3):277-93
8520220
J Mol Biol. 1998 Jul 31;280(5):933-52
9671561
J Cell Sci. 2007 May 1;120(Pt 9):1615-23
17405812
Proteins. 1992 Apr;12(4):345-64
1579569
Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9
2025413
J Biol Chem. 1993 Dec 15;268(35):26571-7
7504673
Methods Mol Biol. 2004;278:353-78
15318003
Biochemistry. 1998 Jul 7;37(27):9784-92
9657692
Cell Mol Life Sci. 2004 Jul;61(13):1546-61
15224180
Neuron. 2003 Oct 9;40(2):427-46
14556719
J Mol Biol. 2008 Mar 14;377(1):162-80
18241885
Biochemistry. 1992 Sep 15;31(36):8571-86
1390643
J Biol Chem. 1978 Sep 25;253(18):6551-60
681366
FASEB J. 2003 Nov;17(14):2014-24
14597671
J Biol Chem. 1996 Nov 15;271(46):28766-71
8910518