Adapted ATPase domain communication overcomes the cytotoxicity of p97 inhibitors.


Abstract

The AAA+ ATPase p97 regulates ubiquitin-dependent protein homeostasis and has been pursued as a cancer drug target. The ATP-competitive inhibitor CB-5083 and allosteric inhibitor NMS-873 are the most advanced p97 inhibitors described to date. Previous studies have reported that their cytotoxicity can be readily overcome and involves single p97 mutations in the linker between the D1 and D2 ATPase domains and within D2. We report here that the proline 472 to leucine (P472L) mutation, in the D1-D2 linker and identified in CB-5083-resistant cells, desensitizes p97 to both inhibitor classes. This mutation does not disrupt the distinct D2-binding sites of the inhibitors. Instead, P472L changes ATPase domain communication within the p97 hexamer. P472L enhances cooperative D2 ATP binding and hydrolysis. This mechanism alters the function of the D1-D2 linker in the control of D2 activity involving the ATP-bound state of D1. Although increased D2 activity is sufficient to desensitize the P472L mutant to NMS-873, the mutant's desensitization to CB-5083 also requires D1 ATPase domain function. Our study highlights the remarkable adaptability of p97 ATPase domain communication that enables escape from mechanistically distinct classes of cytotoxic p97 inhibitors.

Submission Details

ID: btD9qUb7

Submitter: Shu-Ching Ou

Submission Date: March 7, 2019, 11:32 a.m.

Version: 1

Publication Details
Wei Y;Toth JI;Blanco GA;Bobkov AA;Petroski MD,J Biol Chem (2018) Adapted ATPase domain communication overcomes the cytotoxicity of p97 inhibitors. PMID:30381397
Additional Information

ND means not determined.

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1S3S 2004-01-14T00:00:00+0000 2.9 Crystal structure of AAA ATPase p97/VCP ND1 in complex with p47 C
3CF1 2008-03-01T00:00:00+0000 4.4 Structure of P97/vcp in complex with ADP/ADP.alfx
3CF0 2008-03-01T00:00:00+0000 3.0 Structure of D2 subdomain of P97/VCP in complex with ADP
3CF2 2008-03-01T00:00:00+0000 3.5 Structure of P97/vcp in complex with ADP/AMP-PNP
3CF3 2008-03-01T00:00:00+0000 4.25 Structure of P97/vcp in complex with ADP
1E32 2000-06-05T00:00:00+0000 2.9 Structure of the N-Terminal domain and the D1 AAA domain of membrane fusion ATPase p97
1R7R 2003-10-22T00:00:00+0000 3.6 The crystal structure of murine p97/VCP at 3.6A
2PJH 2007-04-16T00:00:00+0000 0 Strctural Model of the p97 N domain- npl4 UBD complex
4KLN 2013-05-07T00:00:00+0000 2.62 Structure of p97 N-D1 A232E mutant in complex with ATPgS
5C1A 2015-06-13T00:00:00+0000 3.8 p97-N750D/R753D/M757D/Q760D in complex with ATP-gamma-S

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
584.4 A,B,C,D,E,F Transitional endoplasmic reticulum ATPase Q7ZU99 TERA_DANRE
573.0 A,B,C,D,E,F Transitional endoplasmic reticulum ATPase P23787 TERA_XENLA
576.0 A,B,C,D,E,F Transitional endoplasmic reticulum ATPase Q6GL04 TERA_XENTR
598.8 A,B,C,D,E,F Transitional endoplasmic reticulum ATPase P03974 TERA_PIG
598.8 A,B,C,D,E,F Transitional endoplasmic reticulum ATPase Q3ZBT1 TERA_BOVIN
599.4 A,B,C,D,E,F Transitional endoplasmic reticulum ATPase P46462 TERA_RAT
600.0 A,B,C,D,E,F Transitional endoplasmic reticulum ATPase Q01853 TERA_MOUSE
600.0 A,B,C,D,E,F Transitional endoplasmic reticulum ATPase P55072 TERA_HUMAN