Sequence-specific recognition of a PxLPxI/L motif by an ankyrin repeat tumbler lock

Chao Xu, Jing Jin, Chuanbing Bian, Robert Lam, Ruijun Tian, Ryan Weist, Linya You, Jianyun Nie, Alexey Bochkarev, Wolfram Tempel, Chris S. Tan, Gregory A. Wasney, Masoud Vedadi, Gerald D. Gish, Cheryl H. Arrowsmith, Tony Pawson, Xiang Jiao Yang, Jinrong Min*

*Corresponding author for this work

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Ankyrin repeat family A protein 2 (ANKRA2) interacts with the plasma membrane receptor megalin and the class IIa histone deacetylases HDAC4 and HDAC5. We report that the ankyrin repeat domains of ANKRA2 and its close paralog regulatory factor X-associated ankyrin-containing protein (RFXANK) recognize a PxLPxI/Lmotif found in diverse binding proteins, including HDAC4, HDAC5, HDAC9,megalin, and regulatory factor X, 5 (RFX5). Crystal structures of the ankyrin repeat domain of ANKRA2 in complex with its binding peptides revealed that each of the middle three ankyrin repeats of ANKRA2 recognizes a residue from the PxLPxI/L motif in a tumbler-lock binding mode, with ANKRA2 acting as the lock and the linear binding motif serving as the key. Structural analysis showed that three diseasecausing mutations in RFXANK affect residues that are critical for binding to RFX5. These results suggest a fundamental principle of longitudinal recognition of linear sequences by a repeat-type domain. In addition, phosphorylation of serine 350, a residue embedded within the PxLPxI/L motif of HDAC4, impaired the binding of ANKRA2 but generated a high-affinity docking site for 14-3-3 proteins, which may help sequester this HDAC in the cytoplasm. Thus, the binding preference of the PxLPxI/L motif is signaldependent. Furthermore, proteome-wide screening suggested that a similar phosphorylation-dependent switch may operate in other pathways. Together, our findings uncover a previously uncharacterized sequence- and signal-dependent peptide recognition mode for a repeat-type protein domain.

Original languageEnglish (US)
Article numberra39
JournalScience Signaling
Volume5
Issue number226
DOIs
StatePublished - May 29 2012

Fingerprint

Ankyrin Repeat
Factor X
Proteins
Low Density Lipoprotein Receptor-Related Protein-2
Ankyrins
Phosphorylation
14-3-3 Proteins
Peptides
Histone Deacetylases
Proteome
Cell membranes
Protein Sorting Signals
Structural analysis
Serine
Carrier Proteins
Screening
Cytoplasm
Crystal structure
Switches
Cell Membrane

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Xu, Chao ; Jin, Jing ; Bian, Chuanbing ; Lam, Robert ; Tian, Ruijun ; Weist, Ryan ; You, Linya ; Nie, Jianyun ; Bochkarev, Alexey ; Tempel, Wolfram ; Tan, Chris S. ; Wasney, Gregory A. ; Vedadi, Masoud ; Gish, Gerald D. ; Arrowsmith, Cheryl H. ; Pawson, Tony ; Yang, Xiang Jiao ; Min, Jinrong. / Sequence-specific recognition of a PxLPxI/L motif by an ankyrin repeat tumbler lock. In: Science Signaling. 2012 ; Vol. 5, No. 226.
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title = "Sequence-specific recognition of a PxLPxI/L motif by an ankyrin repeat tumbler lock",
abstract = "Ankyrin repeat family A protein 2 (ANKRA2) interacts with the plasma membrane receptor megalin and the class IIa histone deacetylases HDAC4 and HDAC5. We report that the ankyrin repeat domains of ANKRA2 and its close paralog regulatory factor X-associated ankyrin-containing protein (RFXANK) recognize a PxLPxI/Lmotif found in diverse binding proteins, including HDAC4, HDAC5, HDAC9,megalin, and regulatory factor X, 5 (RFX5). Crystal structures of the ankyrin repeat domain of ANKRA2 in complex with its binding peptides revealed that each of the middle three ankyrin repeats of ANKRA2 recognizes a residue from the PxLPxI/L motif in a tumbler-lock binding mode, with ANKRA2 acting as the lock and the linear binding motif serving as the key. Structural analysis showed that three diseasecausing mutations in RFXANK affect residues that are critical for binding to RFX5. These results suggest a fundamental principle of longitudinal recognition of linear sequences by a repeat-type domain. In addition, phosphorylation of serine 350, a residue embedded within the PxLPxI/L motif of HDAC4, impaired the binding of ANKRA2 but generated a high-affinity docking site for 14-3-3 proteins, which may help sequester this HDAC in the cytoplasm. Thus, the binding preference of the PxLPxI/L motif is signaldependent. Furthermore, proteome-wide screening suggested that a similar phosphorylation-dependent switch may operate in other pathways. Together, our findings uncover a previously uncharacterized sequence- and signal-dependent peptide recognition mode for a repeat-type protein domain.",
author = "Chao Xu and Jing Jin and Chuanbing Bian and Robert Lam and Ruijun Tian and Ryan Weist and Linya You and Jianyun Nie and Alexey Bochkarev and Wolfram Tempel and Tan, {Chris S.} and Wasney, {Gregory A.} and Masoud Vedadi and Gish, {Gerald D.} and Arrowsmith, {Cheryl H.} and Tony Pawson and Yang, {Xiang Jiao} and Jinrong Min",
year = "2012",
month = "5",
day = "29",
doi = "10.1126/scisignal.2002979",
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Xu, C, Jin, J, Bian, C, Lam, R, Tian, R, Weist, R, You, L, Nie, J, Bochkarev, A, Tempel, W, Tan, CS, Wasney, GA, Vedadi, M, Gish, GD, Arrowsmith, CH, Pawson, T, Yang, XJ & Min, J 2012, 'Sequence-specific recognition of a PxLPxI/L motif by an ankyrin repeat tumbler lock', Science Signaling, vol. 5, no. 226, ra39. https://doi.org/10.1126/scisignal.2002979

Sequence-specific recognition of a PxLPxI/L motif by an ankyrin repeat tumbler lock. / Xu, Chao; Jin, Jing; Bian, Chuanbing; Lam, Robert; Tian, Ruijun; Weist, Ryan; You, Linya; Nie, Jianyun; Bochkarev, Alexey; Tempel, Wolfram; Tan, Chris S.; Wasney, Gregory A.; Vedadi, Masoud; Gish, Gerald D.; Arrowsmith, Cheryl H.; Pawson, Tony; Yang, Xiang Jiao; Min, Jinrong.

In: Science Signaling, Vol. 5, No. 226, ra39, 29.05.2012.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Sequence-specific recognition of a PxLPxI/L motif by an ankyrin repeat tumbler lock

AU - Xu, Chao

AU - Jin, Jing

AU - Bian, Chuanbing

AU - Lam, Robert

AU - Tian, Ruijun

AU - Weist, Ryan

AU - You, Linya

AU - Nie, Jianyun

AU - Bochkarev, Alexey

AU - Tempel, Wolfram

AU - Tan, Chris S.

AU - Wasney, Gregory A.

AU - Vedadi, Masoud

AU - Gish, Gerald D.

AU - Arrowsmith, Cheryl H.

AU - Pawson, Tony

AU - Yang, Xiang Jiao

AU - Min, Jinrong

PY - 2012/5/29

Y1 - 2012/5/29

N2 - Ankyrin repeat family A protein 2 (ANKRA2) interacts with the plasma membrane receptor megalin and the class IIa histone deacetylases HDAC4 and HDAC5. We report that the ankyrin repeat domains of ANKRA2 and its close paralog regulatory factor X-associated ankyrin-containing protein (RFXANK) recognize a PxLPxI/Lmotif found in diverse binding proteins, including HDAC4, HDAC5, HDAC9,megalin, and regulatory factor X, 5 (RFX5). Crystal structures of the ankyrin repeat domain of ANKRA2 in complex with its binding peptides revealed that each of the middle three ankyrin repeats of ANKRA2 recognizes a residue from the PxLPxI/L motif in a tumbler-lock binding mode, with ANKRA2 acting as the lock and the linear binding motif serving as the key. Structural analysis showed that three diseasecausing mutations in RFXANK affect residues that are critical for binding to RFX5. These results suggest a fundamental principle of longitudinal recognition of linear sequences by a repeat-type domain. In addition, phosphorylation of serine 350, a residue embedded within the PxLPxI/L motif of HDAC4, impaired the binding of ANKRA2 but generated a high-affinity docking site for 14-3-3 proteins, which may help sequester this HDAC in the cytoplasm. Thus, the binding preference of the PxLPxI/L motif is signaldependent. Furthermore, proteome-wide screening suggested that a similar phosphorylation-dependent switch may operate in other pathways. Together, our findings uncover a previously uncharacterized sequence- and signal-dependent peptide recognition mode for a repeat-type protein domain.

AB - Ankyrin repeat family A protein 2 (ANKRA2) interacts with the plasma membrane receptor megalin and the class IIa histone deacetylases HDAC4 and HDAC5. We report that the ankyrin repeat domains of ANKRA2 and its close paralog regulatory factor X-associated ankyrin-containing protein (RFXANK) recognize a PxLPxI/Lmotif found in diverse binding proteins, including HDAC4, HDAC5, HDAC9,megalin, and regulatory factor X, 5 (RFX5). Crystal structures of the ankyrin repeat domain of ANKRA2 in complex with its binding peptides revealed that each of the middle three ankyrin repeats of ANKRA2 recognizes a residue from the PxLPxI/L motif in a tumbler-lock binding mode, with ANKRA2 acting as the lock and the linear binding motif serving as the key. Structural analysis showed that three diseasecausing mutations in RFXANK affect residues that are critical for binding to RFX5. These results suggest a fundamental principle of longitudinal recognition of linear sequences by a repeat-type domain. In addition, phosphorylation of serine 350, a residue embedded within the PxLPxI/L motif of HDAC4, impaired the binding of ANKRA2 but generated a high-affinity docking site for 14-3-3 proteins, which may help sequester this HDAC in the cytoplasm. Thus, the binding preference of the PxLPxI/L motif is signaldependent. Furthermore, proteome-wide screening suggested that a similar phosphorylation-dependent switch may operate in other pathways. Together, our findings uncover a previously uncharacterized sequence- and signal-dependent peptide recognition mode for a repeat-type protein domain.

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