The crystal structure of dynein intermediate chain-light chain roadblock complex gives new insights into dynein assembly.
The roadblock/LC7 dynein light chain is a ubiquitous component of all dyneins and is essential for many diverse processes including proper axonal transport and dendrite growth. In addition, LC7 functions in non-dynein transcriptional activation of the transforming growth factor-beta complex. Crystal structures of Drosophila melanogaster LC7 in the apo form and in complex with a segment of the disordered N-terminal domain of dynein intermediate chain (IC) provide the first definitive identification of the IC sequence recognized by LC7. The site, confirmed by isothermal titration calorimetry studies, overlaps the IC sequence considered in the literature to be an IC self-association domain. The IC peptide binds as two amphipathic helices that lie along an extensive hydrophobic cleft on LC7 and ends with a polar side-chain interaction network that includes conserved residues from both proteins. The LC7 recognition sequence on IC and its interface with LC7 are well conserved and are, thus, likely representative of all IC x LC7 structures. Interestingly, the position of bound IC in the IC x LC7 complex mimics a helix that is integrated into the primary structure in distantly related LC7 homologs. The IC x LC7 structure further shows that the naturally occurring robl(Z) deletion mutation contains the majority of the IC binding site and suggests that promotion of IC binding by phosphorylation of LC7 is an indirect effect.
Hall
Justin
J
Department of Biochemistry and Biophysics, Oregon State University, Corvallis, Oregon 97331, USA.
Song
Yujuan
Y
Karplus
P Andrew
PA
Barbar
Elisar
E
D000595:Amino Acid Sequence; D000818:Animals; D001059:Apoproteins / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D002151:Calorimetry; D002352:Carrier Proteins / Q000737:chemistry*; D002850:Chromatography, Gel; D018360:Crystallography, X-Ray; D029721:Drosophila Proteins / Q000737:chemistry*; D004331:Drosophila melanogaster / Q000378:metabolism*; D004398:Dyneins / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D017433:Protein Structure, Secondary; D055672:Static Electricity; D013816:Thermodynamics
D001059:Apoproteins; D002352:Carrier Proteins; D029721:Drosophila Proteins; C120447:robl protein, Drosophila; D004398:Dyneins
Am J Physiol Gastrointest Liver Physiol. 2009 Sep;297(3):G480-7
19571232
Acta Crystallogr D Biol Crystallogr. 2006 Jan;62(Pt 1):72-82
16369096
Mol Cell Biol. 1998 Nov;18(11):6816-25
9774695
Proteins. 2003 Feb 15;50(3):437-50
12557186
Mol Cell. 2000 May;5(5):865-76
10882122
J Neurobiol. 2004 Feb 5;58(2):189-200
14704951
Acta Crystallogr D Biol Crystallogr. 2007 Jan;63(Pt 1):32-41
17164524
Methods Mol Biol. 1999;112:531-52
10027275
Traffic. 2007 Dec;8(12):1708-1721
17897317
Cell Motil Cytoskeleton. 2004 Apr;57(4):233-45
14752807
J Mol Biol. 2008 Dec 26;384(4):954-66
18948118
Annu Rev Biochem. 2006;75:607-27
16756504
Proc Natl Acad Sci U S A. 2007 Jun 12;104(24):10028-33
17551010
J Mol Biol. 2006 Oct 6;362(5):1082-93
16949604
J Biol Chem. 2002 Sep 6;277(36):32939-46
12077152
J Virol. 2000 Feb;74(3):1355-63
10627546
Development. 2003 Mar;130(6):1203-13
12571111
Cancer Res. 2005 Aug 1;65(15):6526-33
16061631
J Biol Chem. 2009 Dec 11;284(50):34760-8
19837669
Nat Cell Biol. 2000 Jan;2(1):20-4
10620802
Biochemistry. 2004 Dec 14;43(49):15595-603
15581372
Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
15299926
J Cell Biol. 1996 Feb;132(4):617-33
8647893
Mol Biol Cell. 2002 Dec;13(12):4484-96
12475967
Hum Reprod. 2008 Aug;23(8):1957-62
18492703
J Mol Biol. 1999 Dec 17;294(5):1351-62
10600390
Mol Biol Cell. 2007 Jun;18(6):2254-63
17429069
Cell Motil Cytoskeleton. 2008 Mar;65(3):183-96
18044744
Biochemistry. 2002 Apr 2;41(13):4302-11
11914076
Biochem Biophys Res Commun. 2006 Oct 27;349(3):1125-9
16970917
J Biol Chem. 2006 Apr 7;281(14):9552-9
16452477
J Cell Physiol. 2009 Dec;221(3):707-15
19711352
J Am Chem Soc. 2003 Dec 3;125(48):14859-66
14640663
J Neurocytol. 2000 Nov-Dec;29(11-12):819-29
11466473
J Cell Biol. 1999 Jul 12;146(1):165-80
10402468
Nat Cell Biol. 2001 Oct;3(10):913-7
11584273
Protein Sci. 2009 Jun;18(6):1321-5
19472372
Acta Crystallogr D Biol Crystallogr. 2006 Jan;62(Pt 1):48-57
16369093
Gene. 2001 Dec 27;281(1-2):103-13
11750132
Nat Struct Biol. 1997 Apr;4(4):269-75
9095194
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
15572765
Trends Biochem Sci. 2003 Jul;28(7):361-8
12878003
J Mol Biol. 2007 Aug 10;371(2):457-68
17570393
J Biol Chem. 2009 Nov 27;284(48):33115-21
19759397
Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5669-74
19293377
Proc Natl Acad Sci U S A. 2006 May 23;103(21):8060-5
16690741
J Comput Chem. 2002 Apr 30;23(6):600-9
11939594
J Mol Biol. 2005 Sep 16;352(2):338-54
16083906
J Virol. 2009 Jan;83(2):673-86
18987134
J Neurosci Res. 2007 Sep;85(12):2557-66
17265455
J Mol Biol. 2005 Dec 16;354(5):1043-51
16289575
J Mol Biol. 2003 May 9;328(4):893-907
12729762
Annu Rev Biochem. 2000;69:95-114
10966454
Curr Biol. 2000 Nov 2;10(21):R774-6
11084347
Methods. 2000 Dec;22(4):285-98
11133235