Location of tyrosine 315, a target for phosphorylation by cAbl tyrosine kinase, at the edge of the subunit-subunit interface of the human Rad51 filament

Sebastien Conilleau, Yoshimasa Takizawa, Hiroaki Tachiwana, Fabrice Fleury, Hitoshi Kurumizaka, Masayuki Takahashi

    Research output: Contribution to journalArticle

    17 Citations (Scopus)

    Abstract

    Rad51 is a key element of recombinational DNA repair and its activity is regulated by phosphorylation of the tyrosine residue at position 315 by cAbl kinase. This phosphorylation could be involved in the resistance of cancer cells to chemotherapy. We have investigated the role of this residue by comparing the three-dimensional structures of human Rad51 and its prokaryotic homologue, Escherichia coli RecA. The residue appeared to be on the edge of the subunit-subunit interacting site. The fluorescence intensity of the tryptophan residue inserted at position 315 of human Rad51 in the place of tyrosine was decreased by adding 3 M urea, although the protein was not unfolded as there was no large change in the fluorescence peak position or circular dichroism signal. This change in fluorescence occurred at a lower urea concentration when the protein was diluted, which favours dissociation. These results indicate that the change is related to the dissociation of Rad51 polymer and that residue 315 is close to the subunit-subunit interacting site. ATP and ADP, which affect the filament structure, caused a blue shift in the fluorescence peak. These nucleotides probably altered the subunit-subunit contacts and may thus affect the filament structure. Phosphorylation of this residue could therefore affect the formation and structure of the Rad51 filament. Correct prediction of subunit-subunit interface of Rad51 by simple comparison of structures of Rad51 and RecA supports the idea that Rad51 forms the filament in a similar way as does RecA.

    Original languageEnglish
    Pages (from-to)797-804
    Number of pages8
    JournalJournal of Molecular Biology
    Volume339
    Issue number4
    DOIs
    Publication statusPublished - 2004 Jun 11

    Fingerprint

    Protein-Tyrosine Kinases
    Tyrosine
    Fluorescence
    Phosphorylation
    Urea
    Recombinational DNA Repair
    Circular Dichroism
    Tryptophan
    Adenosine Diphosphate
    Polymers
    Proteins
    Phosphotransferases
    Nucleotides
    Adenosine Triphosphate
    Escherichia coli
    Drug Therapy
    Neoplasms

    Keywords

    • cAbl tyrosine kinase
    • CD, circular dichroism
    • filament formation
    • HsRad51, human Rad51
    • protein phosphorylation
    • Rad51 protein
    • recombinational repair
    • XlRad51.1, Xenopus Rad51

    ASJC Scopus subject areas

    • Virology

    Cite this

    Location of tyrosine 315, a target for phosphorylation by cAbl tyrosine kinase, at the edge of the subunit-subunit interface of the human Rad51 filament. / Conilleau, Sebastien; Takizawa, Yoshimasa; Tachiwana, Hiroaki; Fleury, Fabrice; Kurumizaka, Hitoshi; Takahashi, Masayuki.

    In: Journal of Molecular Biology, Vol. 339, No. 4, 11.06.2004, p. 797-804.

    Research output: Contribution to journalArticle

    Conilleau, Sebastien ; Takizawa, Yoshimasa ; Tachiwana, Hiroaki ; Fleury, Fabrice ; Kurumizaka, Hitoshi ; Takahashi, Masayuki. / Location of tyrosine 315, a target for phosphorylation by cAbl tyrosine kinase, at the edge of the subunit-subunit interface of the human Rad51 filament. In: Journal of Molecular Biology. 2004 ; Vol. 339, No. 4. pp. 797-804.
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    AU - Takizawa, Yoshimasa

    AU - Tachiwana, Hiroaki

    AU - Fleury, Fabrice

    AU - Kurumizaka, Hitoshi

    AU - Takahashi, Masayuki

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    AB - Rad51 is a key element of recombinational DNA repair and its activity is regulated by phosphorylation of the tyrosine residue at position 315 by cAbl kinase. This phosphorylation could be involved in the resistance of cancer cells to chemotherapy. We have investigated the role of this residue by comparing the three-dimensional structures of human Rad51 and its prokaryotic homologue, Escherichia coli RecA. The residue appeared to be on the edge of the subunit-subunit interacting site. The fluorescence intensity of the tryptophan residue inserted at position 315 of human Rad51 in the place of tyrosine was decreased by adding 3 M urea, although the protein was not unfolded as there was no large change in the fluorescence peak position or circular dichroism signal. This change in fluorescence occurred at a lower urea concentration when the protein was diluted, which favours dissociation. These results indicate that the change is related to the dissociation of Rad51 polymer and that residue 315 is close to the subunit-subunit interacting site. ATP and ADP, which affect the filament structure, caused a blue shift in the fluorescence peak. These nucleotides probably altered the subunit-subunit contacts and may thus affect the filament structure. Phosphorylation of this residue could therefore affect the formation and structure of the Rad51 filament. Correct prediction of subunit-subunit interface of Rad51 by simple comparison of structures of Rad51 and RecA supports the idea that Rad51 forms the filament in a similar way as does RecA.

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