Proteomics

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Identification of HJURP binding determinants on the Mis18 complex using photo-cross-linking and mass spectrometry

ABSTRACT: In the early G1 phase of the cell cycle, human HJURP is recruited to centromeres by the Mis18 complex, which consists of Mis18α, Mis18β and M18BP1. This interaction of HJURP with the Mis18 complex is essential for CENP-A deposition and the preservation of centromere identity. To identify the binding site of HJURP on the Mis18 complex, we used the amber-codon suppression method to incorporate unnatural photo-cross-linking amino acids (Bpa, p-benzoyl-L-phenylalanine; AbK, 3’-azibutyl-N-carbamoyl-lysine) and performed UV-cross-linking and mass spectrometry (MS) experiments.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Dongqing Pan  

LAB HEAD: Andrea Musacchio

PROVIDER: PXD013339 | Pride | 2019-08-01

REPOSITORIES: Pride

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20181124_Q2_U1_1-10_AB_SA_DOP_20181116_HJ2.raw Raw
20181124_Q2_U1_1-10_AB_SA_DOP_20181116_HJ8.raw Raw
20181124_Q2_U1_1-2_AB_SA_DOP_20181116_HJ1.raw Raw
20181124_Q2_U1_1-2_AB_SA_DOP_20181116_HJ4-1.raw Raw
20181124_Q2_U1_1-2_AB_SA_DOP_20181116_HJ6.raw Raw
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Publications

Mechanism of centromere recruitment of the CENP-A chaperone HJURP and its implications for centromere licensing.

Pan Dongqing D   Walstein Kai K   Take Annika A   Bier David D   Kaiser Nadine N   Musacchio Andrea A  

Nature communications 20190906 1

Nucleosomes containing the histone H3 variant CENP-A are the epigenetic mark of centromeres, the kinetochore assembly sites required for chromosome segregation. HJURP is the CENP-A chaperone, which associates with Mis18α, Mis18β, and M18BP1 to target centromeres and deposit new CENP-A. How these proteins interact to promote CENP-A deposition remains poorly understood. Here we show that two repeats in human HJURP proposed to be functionally distinct are in fact interchangeable and bind concomitan  ...[more]

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