A dynamic charge-charge interaction modulates PP2A:B56 substrate recruitment

Abstract
Data availability
Article and author information
Metrics

Abstract

The recruitment of substrates by the ser/thr protein phosphatase 2A (PP2A) is poorly understood, limiting our understanding of PP2A-regulated signaling. Recently, the first PP2A:B56 consensus binding motif, LxxIxE, was identified. However, most validated LxxIxE motifs bind PP2A:B56 with micromolar affinities, suggesting that additional motifs exist to enhance PP2A:B56 binding. Here, we report the requirement of a positively charged motif in a subset of PP2A:B56 interactors, including KIF4A, to facilitate B56 binding via dynamic, electrostatic interactions. Using molecular and cellular experiments, we show that a conserved, negatively charged groove on B56 mediates dynamic binding. We also discovered that this positively charged motif, in addition to facilitating KIF4A dephosphorylation, is essential for condensin I binding, a function distinct and exclusive from PP2A-B56 binding. Together, these results reveal how dynamic, charge-charge interactions fine-tune the interactions mediated by specific motifs, providing a new framework for understanding how PP2A regulation drives cellular signaling.

Data availability

All NMR chemical shifts have been deposited in the BioMagResBank (BMRB 27913). Atomic coordinates and structure factors have been deposited in the Protein Data Bank (6OYL, 6VRO). The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (Vizcaíno et al., 2014) through the PRIDE partner repository (PXD013886).

The following data sets were generated

1. Page R et al
(2020) Atomic coordinates and structure factors
Protein Data Bank, 6OYL.

http://www.rcsb.org/structure/6OYL
1. Page R et al
(2020) Atomic coordinates and structure factors
Protein Data Bank, 6VRO.

http://www.rcsb.org/structure/6VRO
1. Wang X
2. Garvanska D
3. Kettenbach A
4. Peti W
5. Nilsson J
6. Page R
(2020) Backbone 1H, 13C, and 15N Chemical Shift Assignments for the C-terminal Fragment of a Kinesin KIF4A Variant
Biological Magnetic Resonance Data Bank, 27913.

http://bmrb.wisc.edu/data_library/summary/index.php?bmrbId=27913
1. Page R et al
(2020) Mass spectrometry proteomics data
Proteome Exchange, PXD013886.

http://proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD013886

The following previously published data sets were used

1. Kettenbach A
(2019) A dynamic charge:charge interaction modulates PP2A:B56 interactions
MassIVE, MSV000083785.

https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=9a6d3fdf30db4f6db9243d3d3cc07058

Article and author information

Author details

Xinru Wang

Department of Chemistry and Biochemistry, University of Arizona, Tucson, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-5994-707X
Dimitriya H Garvanska

The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark

Competing interests
The authors declare that no competing interests exist.
Isha Nasa

Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth, Lebanon, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-7699-795X
Yumi Ueki

The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark

Competing interests
The authors declare that no competing interests exist.
Gang Zhang

The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-7697-7203
Arminja N Kettenbach

Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth, Lebanon, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0003-3979-4576
Wolfgang Peti

Department of Chemistry and Biochemistry, University of Arizona, Tucson, United States

Competing interests
The authors declare that no competing interests exist.
Jakob Nilsson

The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Germany

For correspondence
jakob.nilsson@cpr.ku.dk

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0003-4100-1125
Rebecca Page

Department of Chemistry and Biochemistry, University of Arizona, Tucson, United States

For correspondence
rebeccapage@email.arizona.edu

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-4645-1232

Funding

National Institute of General Medical Sciences (R35GM119455)

Arminja N Kettenbach

National Institute of General Medical Sciences (P20GM113132)

Arminja N Kettenbach

National Institute of General Medical Sciences (R01GM098482)

Rebecca Page

National Institute of Neurological Disorders and Stroke (R01NS091336)

Wolfgang Peti

National Institute of General Medical Sciences (R01GM134683)

Wolfgang Peti

Novo Nordisk (NNF14CC0001)

Jakob Nilsson

Independent Research Fund Denmark (DFF-7016-00086)

Jakob Nilsson

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.