LIG_APCC_Dbox_1
ELM server details
ELM
blue dot
Functional site class:
APCC-binding Destruction motifs
Functional site description:
The anaphase-promoting ubiquitin ligase complex APC/C selectively targets numerous cell cycle-regulatory proteins for ubiquitin-mediated proteasome-dependent degradation. The targets of the APC/C are degraded in an ordered, sequential manner which ensures the correct progression of the cell cycle. Cdh1 and Cdc20 are WD-repeat containing proteins which act as co-activators of the APC/C at distinct steps of the cycle. Cdc20 joins the APC/C in early mitosis and is then replaced by Cdh1 during anaphase. Both Cdh1 and Cdc20 recognize the target proteins via short, very specific "destruction motifs". The motifs allow recruitment of the targets, to the APC/C complex which subsequently poly-ubiquitinates them.
ELM(s): LIG_APCC_KENbox_2, LIG_APCC_Dbox_1
LIG_APCC_Dbox_1 description: The Destruction Box is a short sequence motif, comprising a highly conserved RXXL motif. It is found in several key cell cycle proteins where it acts as a signal for cycle-dependent proteolysis. It was originally found in the cyclin B protein (Glotzer et al., 1991, Nature 349:132-8). The motif is necessary to induce cyclin B degradation, and when fused to a foreign protein it confers a cycle-dependent proteolytic pattern similar to that observed for cyclin B. Subsequent studies demonstrated a Dbox-dependent degradation of other key cell-cycle players such as cyclin A, Nek2, Aurora A, Aurora B, securin and geminin (reviewed by Castro et al., 2005, Oncogene 24:314-25).The Dbox is specifically recognized by two proteins belonging to the seven-blade propeller WD40 family: Cdc20 and Cdh1/Hct. Dbox recognition by either Cdc20 or Cdh1/Hct1, stimulates recruitment of the Dbox-containing protein to the anaphase-promoting ubiquitin ligase complex (APC/C). This activates the APC/C ligase to poly-ubiquitinate the Dbox-containing protein, thereby targetting it for destruction. Cdc20 exclusively binds Dbox-containing substrates, whereas Cdh1/Hct1 can interact with substrates containing a Dbox and/or a KEN-box (an additional cell-cycle regulatory degradation motif).
Pattern: .R..L.
Present in taxon(s): Eukaryota  
Not represented in taxon(s):

o See instances for LIG_APCC_Dbox_1


o Abstract

Progress of cell division is governed by the sequential degradation, mediated by the ubiquitination pathway, of proteins playing a key role in the cell cycle. There are two E3 ubiquitin protein ligase complexes which play a role in the cell cycle: the SCF (Skp1/Cullin/F-box) complex and the anaphase-promoting complex (APC/C) (Peters 2006, Castro et al. 2005). The APC/C complex contains at least a dozen different subunits, but it can ubiquitylate substrates only in the presence of the co-activator proteins Cdc20 (also known as fizzy) or Cdh1(also called fzr or Hct1). Cdh1 and Cdc20 are WD40-repeat proteins (well known linear motif-binding domains, folded as 7-blade beta-propellers), which recognise their target proteins via short "degradation motifs". Cdc20 and Cdh1 act at distinct phases of the cell cycle. Early in mitosis, during the metaphase-anaphase transition, APC/Cdc20 is mostly active, whereas Cdh1 is present, but as a phosphorylated inactive form which cannot bind to the APC/C. Later, in mitotic exit and further during the G1 phases, Cdh1 is activated by dephosphorylation, and binds to the APC/C by replacing Cdc20 and promoting its rapid degradation.
A number of degradation motifs have been identified within APC/C substrate proteins. These motifs are generally defined as short conserved sequences whose deletion or mutation promotes the stabilisation of the proteins where they are naturally found, and which can confer cell-cycle dependent degradation on unrelated proteins. The best characterised ones are the Destruction box (D-box) and KEN box.
The D-box was originally found in the cyclin B protein (Glotzer et al., 1991). It is one of the earlier described linear motifs. This short sequence, comprising a highly conserved RXXL motif, is necessary to induce cyclin B degradation. Moreover, when fused to a foreign protein it is sufficient to generate a cycle-dependent proteolytic pattern similar to that observed for cyclin B. Subsequent studies demonstrated a D-box-dependent degradation of other key cell-cycle players such as cyclin A (Lorca et al., 1992), Nek2 (Hames et al., 2001), Aurora A (Castro et al., 2002), Aurora B (Stewart and Fang, 2005), securin (Zou et al., 1999) and geminin (Mc Garry and Kirchner, 1998).
The first KEN box (consensus KEN) was identified within Cdc20 itself (Pfleger and Kirschner 2000). Later, active KEN boxes were also found within human CDC6, securin, Drosophila cyclin A, yeast Hsl1, Clb2, Aurora kinase B, BUB1 and CIN8 (see attached references).
Both D-box and KEN-box are recognised by Cdh1 and/or Cdc20, which subsequently recruit the APC/C complex, leading to the ubiquitination and proteasome-mediated degradation of the target protein.
The D-box is recognized by both Cdc20 and Cdh1, whereas the KEN-box is preferentially recognized by Cdh1. Cdc20 itself contains a KEN box, which is therefore recognized by Cdh1, ensuring the temporal degradation of Cdc20 and its replacement by Cdh1 as a cofactor of the APC/C.
[Experimental studies of APC-target proteins have shown that some of them contain only D-box, others contain only KEN-box, some contain both. D-box and KEN can act as an independent entity or as a co-ordinate unit for protein degradation.
Finally, the presence of D-box or KEN-box motifs in a sequence does not always guarantee that they are active degradation signals for the proteins in which they are found. Indeed, there is always a possibility that the protein in which a potential destruction motif has been mutated becomes resistant to proteasome degradation due to serious misfolding and aggregation, and not because of losing a specific APC/C-targeting site. Putative destruction motifs found within areas predicted to be natively disordered are much more likely to be active than those found in known or predicted globular regions/domains.]

o Selected references

Arlot-Bonnemains Y, Klotzbucher A, Giet R, Uzbekov R, Bihan R, Prigent C
Identification of a functional destruction box in the Xenopus laevis aurora-A kinase pEg2.
FEBS Lett 2001 Nov 9;508(1) : 149-52.
PMID: 11707286

Castro A, Bernis C, Vigneron S, Labbe JC, Lorca T
The anaphase-promoting complex: a key factor in the regulation of cell cycle.
Oncogene 2005 Jan 13;24(3) : 314-25.
PMID: 15678131

Glotzer M, Murray AW, Kirschner MW
Cyclin is degraded by the ubiquitin pathway.
Nature 1991 Jan 10;349(6305) : 132-8.
PMID: 1846030

Hames RS, Wattam SL, Yamano H, Bacchieri R, Fry AM
APC/C-mediated destruction of the centrosomal kinase Nek2A occurs in early mitosis and depends upon a cyclin A-type D-box.
EMBO J 2001 Dec 17;20(24) : 7117-27.
PMID: 11742988

Lindon C, Pines J
Ordered proteolysis in anaphase inactivates Plk1 to contribute to proper mitotic exit in human cells.
J Cell Biol 2004 Jan 19;164(2) : 233-41.
PMID: 14734534

Lorca T, Devault A, Colas P, Van Loon A, Fesquet D, Lazaro JB, Doree M
Cyclin A-Cys41 does not undergo cell cycle-dependent degradation in Xenopus extracts.
FEBS Lett 1992 Jul 13;306(1) : 90-3.
PMID: 1321060

McGarry TJ, Kirschner MW
Geminin, an inhibitor of DNA replication, is degraded during mitosis.
Cell 1998 Jun 12;93(6) : 1043-53.
PMID: 9635433

Nguyen HG, Chinnappan D, Urano T, Ravid K
Mechanism of Aurora-B degradation and its dependency on intact KEN and A-boxes: identification of an aneuploidy-promoting property.
Mol Cell Biol 2005 Jun;25(12) : 4977-92.
PMID: 15923616

Peters JM
The anaphase promoting complex/cyclosome: a machine designed to destroy.
Nat Rev Mol Cell Biol 2006 Sep;7(9) : 644-56.
PMID: 16896351

Petersen BO, Wagener C, Marinoni F, Kramer ER, Melixetian M, Lazzerini Denchi E, Gieffers C, Matteucci C, Peters JM, Helin K
Cell cycle- and cell growth-regulated proteolysis of mammalian CDC6 is dependent on APC-CDH1.
Genes Dev 2000 Sep 15;14(18) : 2330-43.
PMID: 10995389

Stewart S, Fang G
Destruction box-dependent degradation of aurora B is mediated by the anaphase-promoting complex/cyclosome and Cdh1.
Cancer Res 2005 Oct 1;65(19) : 8730-5.
PMID: 16204042

Zou H, McGarry TJ, Bernal T, Kirschner MW
Identification of a vertebrate sister-chromatid separation inhibitor involved in transformation and tumorigenesis.
Science 1999 Jul 16;285(5426) : 418-22.
PMID: 10411507

o This ELM has been assigned the following Gene Ontology (GO) terms for biological process, cellular component and molecular function.

Biological Process
  proteasomal ubiquitin-dependent protein catabolic process
  cell cycle control
Cellular Component
  nucleus
  cytosol
Molecular Function
  protein domain specific binding

 

o Instances for LIG_APCC_Dbox_1

SequencePositionSubsequence
(Click for evidence information)
PDBGene NameProtein DescriptionOrganism
AURKB_HUMAN 314-319 RHNPSERLPLAQVSAHP - Name=AURKB; Synonyms=AIK2, AIM1, ARK2, STK12; Serine/threonine-protein kinase 12 (EC 2.7.11.1) (Aurora-B) (Aurora- and Ipl1-like midbody-associated protein 1) (AIM-1) (Aurora/IPL1- related kinase 2) (Aurora-related kinase 2) (STK-1). Homo sapiens (Human).
AURKB_RAT 317-322 KHNPSQRLPLEQVSAHP - Name=Aurkb; Synonyms=Aim1, Stk12; Serine/threonine-protein kinase 12 (EC 2.7.11.1) (Aurora-B) (Aurora- and Ipl1-like midbody-associated protein 1) (AIM-1). Rattus norvegicus (Rat).
Q9IA80_XENLA 54-59 MVSKPSRKALGNVNKQI - - Securin. Xenopus laevis (African clawed frog).
O93355_XENLA 32-37 NEALAPRRTLKVIQQSA - - Geminin L. Xenopus laevis (African clawed frog).
PLK1_HUMAN 336-341 SLDPSNRKPLTVLNKGL - Name=PLK1; Synonyms=PLK; Serine/threonine-protein kinase PLK1 (EC 2.7.11.21) (Polo-like kinase 1) (PLK-1) (Serine/threonine-protein kinase 13) (STPK13). Homo sapiens (Human).
CDC6_HUMAN 55-60 ALPLSPRKRLGDDNLCN - Name=CDC6; Synonyms=CDC18L; Cell division control protein 6 homolog (CDC6-related protein) (p62(cdc6)) (HsCDC6) (HsCDC18). Homo sapiens (Human).
STK6_HUMAN 370-375 KHNPSQRPMLREVLEHP - Name=AURKA; Synonyms=AIK, ARK1, AURA, BTAK, STK15, STK6; Serine/threonine-protein kinase 6 (EC 2.7.11.1) (Aurora kinase A) (Aurora-A) (Serine/threonine kinase 15) (Aurora/IPL1-related kinase 1) (Aurora-related kinase 1) (hARK1) (Breast tumor-amplified kinase). Homo sapiens (Human).
Q5I1Z9_HUMAN 145-150 YSLLKERKFLSLASNPV - Name=NEK2A; Never in mitosis gene a-related kinase 2A splice variant NEK2A-T (Fragment). Homo sapiens (Human).
ID2_HUMAN 99-104 GQNQASRTPLTTLNTDI - Name=ID2; DNA-binding protein inhibitor ID-2 (Inhibitor of DNA binding 2). Homo sapiens (Human).
CCNB_ARBPU 41-46 VQKPAQRAALGNISNVV - - G2/mitotic-specific cyclin-B. Arbacia punctulata (Punctuate sea urchin).
CCNA1_XENLA 40-45 QPNLPQRTVLGVIGDNE - Name=ccna1; Cyclin-A1. Xenopus laevis (African clawed frog).
STK6_XENLA 377-382 KHNPNHRLPLKGVLEHP - Name=aurka; Synonyms=eg2, stk6; Serine/threonine-protein kinase 6 (EC 2.7.11.1) (Aurora kinase A) (Aurora-A) (Serine/threonine-protein kinase Eg2) (pEg2) (p46Eg265). Xenopus laevis (African clawed frog).