TY - JOUR
T1 - Multivalent coiled-coil interactions enable full-scale centrosome assembly and strength
AU - Rios, Manolo U.
AU - Bagnucka, Małgorzata A.
AU - Ryder, Bryan D.
AU - Ferreira Gomes, Beatriz
AU - Familiari, Nicole E.
AU - Yaguchi, Kan
AU - Amato, Matthew
AU - Stachera, Weronika E.
AU - Joachimiak, Łukasz A.
AU - Woodruff, Jeffrey B.
N1 - Publisher Copyright:
© 2024 Rios et al.
PY - 2024/4/1
Y1 - 2024/4/1
N2 - The outermost layer of centrosomes, called pericentriolar material (PCM), organizes microtubules for mitotic spindle assembly. The molecular interactions that enable PCM to assemble and resist external forces are poorly understood. Here, we use crosslinking mass spectrometry (XL-MS) to analyze PLK-1-potentiated multimerization of SPD-5, the main PCM scaffold protein in C. elegans. In the unassembled state, SPD-5 exhibits numerous intramolecular crosslinks that are eliminated after phosphorylation by PLK-1. Thus, phosphorylation induces a structural opening of SPD-5 that primes it for assembly. Multimerization of SPD-5 is driven by interactions between multiple dispersed coiled-coil domains. Structural analyses of a phosphorylated region (PReM) in SPD-5 revealed a helical hairpin that dimerizes to form a tetrameric coiled-coil. Mutations within this structure and other interacting regions cause PCM assembly defects that are partly rescued by eliminating microtubule-mediated forces, revealing that PCM assembly and strength are interdependent. We propose that PCM size and strength emerge from specific, multivalent coiled-coil interactions between SPD-5 proteins.
AB - The outermost layer of centrosomes, called pericentriolar material (PCM), organizes microtubules for mitotic spindle assembly. The molecular interactions that enable PCM to assemble and resist external forces are poorly understood. Here, we use crosslinking mass spectrometry (XL-MS) to analyze PLK-1-potentiated multimerization of SPD-5, the main PCM scaffold protein in C. elegans. In the unassembled state, SPD-5 exhibits numerous intramolecular crosslinks that are eliminated after phosphorylation by PLK-1. Thus, phosphorylation induces a structural opening of SPD-5 that primes it for assembly. Multimerization of SPD-5 is driven by interactions between multiple dispersed coiled-coil domains. Structural analyses of a phosphorylated region (PReM) in SPD-5 revealed a helical hairpin that dimerizes to form a tetrameric coiled-coil. Mutations within this structure and other interacting regions cause PCM assembly defects that are partly rescued by eliminating microtubule-mediated forces, revealing that PCM assembly and strength are interdependent. We propose that PCM size and strength emerge from specific, multivalent coiled-coil interactions between SPD-5 proteins.
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U2 - 10.1083/jcb.202306142
DO - 10.1083/jcb.202306142
M3 - Article
C2 - 38456967
AN - SCOPUS:85187500055
SN - 0021-9525
VL - 223
JO - The Journal of cell biology
JF - The Journal of cell biology
IS - 4
ER -