Ancient origins of allosteric activation in a Ser-Thr kinase

Science ◽  
2020 ◽  
Vol 367 (6480) ◽  
pp. 912-917 ◽  
Author(s):  
Adelajda Hadzipasic ◽  
Christopher Wilson ◽  
Vy Nguyen ◽  
Nadja Kern ◽  
Chansik Kim ◽  
...  
Keyword(s):  
Activation Mechanism ◽  
Activation Loop ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Allosteric Activation ◽  
Fundamental Interest ◽  
Evolutionary Path ◽  
Cellular Events ◽  
Sequence Reconstruction ◽  
Allosteric Activator

A myriad of cellular events are regulated by allostery; therefore, evolution of this process is of fundamental interest. Here, we use ancestral sequence reconstruction to resurrect ancestors of two colocalizing proteins, Aurora A kinase and its allosteric activator TPX2 (targeting protein for Xklp2), to experimentally characterize the evolutionary path of allosteric activation. Autophosphorylation of the activation loop is the most ancient activation mechanism; it is fully developed in the oldest kinase ancestor and has remained stable over 1 billion years of evolution. As the microtubule-associated protein TPX2 appeared, efficient kinase binding to TPX2 evolved, likely owing to increased fitness by virtue of colocalization. Subsequently, TPX2-mediated allosteric kinase regulation gradually evolved. Surprisingly, evolution of this regulation is encoded in the kinase and did not arise by a dominating mechanism of coevolution.

scholarly journals Detection of Ligand‐induced Conformational Changes in the Activation Loop of Aurora‐A Kinase by PELDOR Spectroscopy

ChemistryOpen ◽  
2016 ◽  
Vol 5 (6) ◽  
pp. 531-534 ◽  
Author(s):  
Selena G. Burgess ◽  
Maria Grazia Concilio ◽  
Richard Bayliss ◽  
Alistair J. Fielding
Keyword(s):  
Conformational Changes ◽  
Activation Loop ◽  
Aurora A Kinase ◽  
Aurora A

scholarly journals Dynamic Equilibrium of the Aurora A Kinase Activation Loop Revealed by Single-Molecule Spectroscopy

2017 ◽  
Vol 56 (38) ◽  
pp. 11409-11414 ◽  
Author(s):  
James A. H. Gilburt ◽  
Hajrah Sarkar ◽  
Peter Sheldrake ◽  
Julian Blagg ◽  
Liming Ying ◽  
...  
Keyword(s):  
Single Molecule ◽  
Dynamic Equilibrium ◽  
Activation Loop ◽  
Single Molecule Spectroscopy ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Kinase Activation

scholarly journals Author response: Molecular mechanism of Aurora A kinase autophosphorylation and its allosteric activation by TPX2

2014 ◽  
Author(s):  
Adelajda Zorba ◽  
Vanessa Buosi ◽  
Steffen Kutter ◽  
Nadja Kern ◽  
Francesco Pontiggia ◽  
...  
Keyword(s):  
Molecular Mechanism ◽  
Author Response ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Allosteric Activation

scholarly journals Dynamic Equilibrium of the Aurora A Kinase Activation Loop Revealed by Single-Molecule Spectroscopy

2017 ◽  
Vol 129 (38) ◽  
pp. 11567-11572
Author(s):  
James A. H. Gilburt ◽  
Hajrah Sarkar ◽  
Peter Sheldrake ◽  
Julian Blagg ◽  
Liming Ying ◽  
...  
Keyword(s):  
Single Molecule ◽  
Dynamic Equilibrium ◽  
Activation Loop ◽  
Single Molecule Spectroscopy ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Kinase Activation

scholarly journals Bora phosphorylation substitutes in trans for T-loop phosphorylation in Aurora A to promote mitotic entry

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
N. Tavernier ◽  
Y. Thomas ◽  
S. Vigneron ◽  
P. Maisonneuve ◽  
S. Orlicky ◽  
...  
Keyword(s):  
Kinase Domain ◽  
Activation Mechanism ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Mitotic Entry ◽  
Amino Acid Region ◽  
Egg Extracts ◽  
Activation Segment ◽  
Xenopus Egg ◽  
Xenopus Egg Extracts

AbstractPolo-like kinase 1 (Plk1) is instrumental for mitotic entry and progression. Plk1 is activated by phosphorylation on a conserved residue Thr210 in its activation segment by the Aurora A kinase (AURKA), a reaction that critically requires the co-factor Bora phosphorylated by a CyclinA/B-Cdk1 kinase. Here we show that phospho-Bora is a direct activator of AURKA kinase activity. We localize the key determinants of phospho-Bora function to a 100 amino acid region encompassing two short Tpx2-like motifs and a phosphoSerine-Proline motif at Serine 112, through which Bora binds AURKA. The latter substitutes in trans for the Thr288 phospho-regulatory site of AURKA, which is essential for an active conformation of the kinase domain. We demonstrate the importance of these determinants for Bora function in mitotic entry both in Xenopus egg extracts and in human cells. Our findings unveil the activation mechanism of AURKA that is critical for mitotic entry.

scholarly journals Molecular mechanism of Aurora A kinase autophosphorylation and its allosteric activation by TPX2

eLife ◽  
2014 ◽  
Vol 3 ◽  
Author(s):  
Adelajda Zorba ◽  
Vanessa Buosi ◽  
Steffen Kutter ◽  
Nadja Kern ◽  
Francesco Pontiggia ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Regulatory Mechanism ◽  
Aurora A Kinase ◽  
Active Conformation ◽  
Aurora A ◽  
Allosteric Activation ◽  
X Ray ◽  
X Ray Crystallography ◽  
Functional Assays ◽  
Spindle Microtubules

We elucidate the molecular mechanisms of two distinct activation strategies (autophosphorylation and TPX2-mediated activation) in human Aurora A kinase. Classic allosteric activation is in play where either activation loop phosphorylation or TPX2 binding to a conserved hydrophobic groove shifts the equilibrium far towards the active conformation. We resolve the controversy about the mechanism of autophosphorylation by demonstrating intermolecular autophosphorylation in a long-lived dimer by combining X-ray crystallography with functional assays. We then address the allosteric activation by TPX2 through activity assays and the crystal structure of a domain-swapped dimer of dephosphorylated Aurora A and TPX21−25. While autophosphorylation is the key regulatory mechanism in the centrosomes in the early stages of mitosis, allosteric activation by TPX2 of dephosphorylated Aurora A could be at play in the spindle microtubules. The mechanistic insights into autophosphorylation and allosteric activation by TPX2 binding proposed here, may have implications for understanding regulation of other protein kinases.

scholarly journals Ligand discrimination between active and inactive activation loop conformations of Aurora-A kinase is unmodified by phosphorylation

2019 ◽  
Vol 10 (14) ◽  
pp. 4069-4076 ◽  
Author(s):  
James A. H. Gilburt ◽  
Paul Girvan ◽  
Julian Blagg ◽  
Liming Ying ◽  
Charlotte A. Dodson
Keyword(s):  
Activation Loop ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Conformational Preference ◽  
Loop Conformations

Activation loop phosphorylation changes the position of equilibrium between DFG-in-like and DFG-out-like conformations but not the conformational preference of inhibitors.

Centrosome Aurora A gradient ensures single polarity axis in C. elegans embryos

2020 ◽  
Vol 48 (3) ◽  
pp. 1243-1253 ◽  
Author(s):  
Sukriti Kapoor ◽  
Sachin Kotak
Keyword(s):  
Symmetry Breaking ◽  
Cell Fate ◽  
Cell Cortex ◽  
Axis Formation ◽  
Aurora A Kinase ◽  
Aurora A ◽  
C Elegans ◽  
Cell Embryo ◽  
Polarity Establishment

Cellular asymmetries are vital for generating cell fate diversity during development and in stem cells. In the newly fertilized Caenorhabditis elegans embryo, centrosomes are responsible for polarity establishment, i.e. anterior–posterior body axis formation. The signal for polarity originates from the centrosomes and is transmitted to the cell cortex, where it disassembles the actomyosin network. This event leads to symmetry breaking and the establishment of distinct domains of evolutionarily conserved PAR proteins. However, the identity of an essential component that localizes to the centrosomes and promotes symmetry breaking was unknown. Recent work has uncovered that the loss of Aurora A kinase (AIR-1 in C. elegans and hereafter referred to as Aurora A) in the one-cell embryo disrupts stereotypical actomyosin-based cortical flows that occur at the time of polarity establishment. This misregulation of actomyosin flow dynamics results in the occurrence of two polarity axes. Notably, the role of Aurora A in ensuring a single polarity axis is independent of its well-established function in centrosome maturation. The mechanism by which Aurora A directs symmetry breaking is likely through direct regulation of Rho-dependent contractility. In this mini-review, we will discuss the unconventional role of Aurora A kinase in polarity establishment in C. elegans embryos and propose a refined model of centrosome-dependent symmetry breaking.

Sign in / Sign up

Export Citation Format

Share Document