The KEN box: an APC recognition signal distinct from the D box targeted by Cdh1

AUTOR(ES)

Pfleger, Cathie M.

FONTE

Cold Spring Harbor Laboratory Press

RESUMO

The ordered progression through the cell cycle depends on regulating the abundance of several proteins through ubiquitin-mediated proteolysis. Degradation is precisely timed and specific. One key component of the degradation system, the anaphase promoting complex (APC), is a ubiquitin protein ligase. It is activated both during mitosis and late in mitosis/G1, by the WD repeat proteins Cdc20 and Cdh1, respectively. These activators target distinct sets of substrates. Cdc20–APC requires a well-defined destruction box (D box), whereas Cdh1–APC confers a different and as yet unidentified specificity. We have determined the sequence specificity for Cdh1–APC using two assays, ubiquitination in a completely defined and purified system and degradation promoted by Cdh1–APC in Xenopus extracts. Cdc20 is itself a Cdh1–APC substrate. Vertebrate Cdc20 lacks a D box and therefore is recognized by Cdh1–APC through a different sequence. By analysis of Cdc20 as a substrate, we have identified a new recognition signal. This signal, composed of K-E-N, serves as a general targeting signal for Cdh1–APC. Like the D box, it is transposable to other proteins. Using the KEN box as a template, we have identified cell cycle genes Nek2 and B99 as additional Cdh1–APC substrates. Mutation in the KEN box stabilizes all three proteins against ubiquitination and degradation.

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