Loss of Hda activity stimulates replication initiation from I-box, but not R4 mutant origins in Escherichia coli

Leise Riber, K. Fujimitsu, T. Katayama, Anders Løbner-Olesen

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    Resumé

    Initiation of chromosome replication in Escherichia coli is limited by the initiator protein DnaA associated with ATP. Within the replication origin, binding sites for DnaA associated with ATP or ADP (R boxes) and the DnaA(ATP) specific sites (I-boxes, tau-boxes and 6-mer sites) are found. We analysed chromosome replication of cells carrying mutations in conserved regions of oriC. Cells carrying mutations in DnaA-boxes I2, I3, R2, R3 and R5 as well as FIS and IHF binding sites resembled wild-type cells with respect to origin concentration. Initiation of replication in these mutants occurred in synchrony or with slight asynchrony only. Furthermore, lack of Hda stimulated initiation in all these mutants. The DnaA(ATP) containing complex that leads to initiation can therefore be formed in the absence of several of the origin DnaA binding sites including both DnaA(ATP) specific I-boxes. However, competition between I-box mutant and wildtype origins, revealed a positive role of I-boxes on initiation. On the other hand, mutations affecting DnaA-box R4 were found to be compromised for initiation and could not be augmented by an increase in cellular DnaA(ATP)/DnaA(ADP) ratio. Compared with the sites tested here, R4 therefore seems to contribute to initiation most critically.
    OriginalsprogEngelsk
    TidsskriftMolecular Microbiology
    Vol/bind71
    Udgave nummer1
    Sider (fra-til)107-122
    ISSN0950-382X
    DOI
    StatusUdgivet - 2009

    Citer dette

    Riber, Leise ; Fujimitsu, K. ; Katayama, T. ; Løbner-Olesen, Anders. / Loss of Hda activity stimulates replication initiation from I-box, but not R4 mutant origins in Escherichia coli. I: Molecular Microbiology. 2009 ; Bind 71, Nr. 1. s. 107-122.
    @article{27cb4bc0fa1711de89af000ea68e967b,
    title = "Loss of Hda activity stimulates replication initiation from I-box, but not R4 mutant origins in Escherichia coli",
    abstract = "Initiation of chromosome replication in Escherichia coli is limited by the initiator protein DnaA associated with ATP. Within the replication origin, binding sites for DnaA associated with ATP or ADP (R boxes) and the DnaA(ATP) specific sites (I-boxes, tau-boxes and 6-mer sites) are found. We analysed chromosome replication of cells carrying mutations in conserved regions of oriC. Cells carrying mutations in DnaA-boxes I2, I3, R2, R3 and R5 as well as FIS and IHF binding sites resembled wild-type cells with respect to origin concentration. Initiation of replication in these mutants occurred in synchrony or with slight asynchrony only. Furthermore, lack of Hda stimulated initiation in all these mutants. The DnaA(ATP) containing complex that leads to initiation can therefore be formed in the absence of several of the origin DnaA binding sites including both DnaA(ATP) specific I-boxes. However, competition between I-box mutant and wildtype origins, revealed a positive role of I-boxes on initiation. On the other hand, mutations affecting DnaA-box R4 were found to be compromised for initiation and could not be augmented by an increase in cellular DnaA(ATP)/DnaA(ADP) ratio. Compared with the sites tested here, R4 therefore seems to contribute to initiation most critically.",
    author = "Leise Riber and K. Fujimitsu and T. Katayama and Anders L{\o}bner-Olesen",
    year = "2009",
    doi = "10.1111/j.1365-2958.2008.06516.x",
    language = "English",
    volume = "71",
    pages = "107--122",
    journal = "Molecular Microbiology",
    issn = "0950-382X",
    publisher = "Wiley-Blackwell Publishing Ltd.",
    number = "1",

    }

    Loss of Hda activity stimulates replication initiation from I-box, but not R4 mutant origins in Escherichia coli. / Riber, Leise; Fujimitsu, K.; Katayama, T.; Løbner-Olesen, Anders.

    I: Molecular Microbiology, Bind 71, Nr. 1, 2009, s. 107-122.

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    TY - JOUR

    T1 - Loss of Hda activity stimulates replication initiation from I-box, but not R4 mutant origins in Escherichia coli

    AU - Riber, Leise

    AU - Fujimitsu, K.

    AU - Katayama, T.

    AU - Løbner-Olesen, Anders

    PY - 2009

    Y1 - 2009

    N2 - Initiation of chromosome replication in Escherichia coli is limited by the initiator protein DnaA associated with ATP. Within the replication origin, binding sites for DnaA associated with ATP or ADP (R boxes) and the DnaA(ATP) specific sites (I-boxes, tau-boxes and 6-mer sites) are found. We analysed chromosome replication of cells carrying mutations in conserved regions of oriC. Cells carrying mutations in DnaA-boxes I2, I3, R2, R3 and R5 as well as FIS and IHF binding sites resembled wild-type cells with respect to origin concentration. Initiation of replication in these mutants occurred in synchrony or with slight asynchrony only. Furthermore, lack of Hda stimulated initiation in all these mutants. The DnaA(ATP) containing complex that leads to initiation can therefore be formed in the absence of several of the origin DnaA binding sites including both DnaA(ATP) specific I-boxes. However, competition between I-box mutant and wildtype origins, revealed a positive role of I-boxes on initiation. On the other hand, mutations affecting DnaA-box R4 were found to be compromised for initiation and could not be augmented by an increase in cellular DnaA(ATP)/DnaA(ADP) ratio. Compared with the sites tested here, R4 therefore seems to contribute to initiation most critically.

    AB - Initiation of chromosome replication in Escherichia coli is limited by the initiator protein DnaA associated with ATP. Within the replication origin, binding sites for DnaA associated with ATP or ADP (R boxes) and the DnaA(ATP) specific sites (I-boxes, tau-boxes and 6-mer sites) are found. We analysed chromosome replication of cells carrying mutations in conserved regions of oriC. Cells carrying mutations in DnaA-boxes I2, I3, R2, R3 and R5 as well as FIS and IHF binding sites resembled wild-type cells with respect to origin concentration. Initiation of replication in these mutants occurred in synchrony or with slight asynchrony only. Furthermore, lack of Hda stimulated initiation in all these mutants. The DnaA(ATP) containing complex that leads to initiation can therefore be formed in the absence of several of the origin DnaA binding sites including both DnaA(ATP) specific I-boxes. However, competition between I-box mutant and wildtype origins, revealed a positive role of I-boxes on initiation. On the other hand, mutations affecting DnaA-box R4 were found to be compromised for initiation and could not be augmented by an increase in cellular DnaA(ATP)/DnaA(ADP) ratio. Compared with the sites tested here, R4 therefore seems to contribute to initiation most critically.

    U2 - 10.1111/j.1365-2958.2008.06516.x

    DO - 10.1111/j.1365-2958.2008.06516.x

    M3 - Journal article

    VL - 71

    SP - 107

    EP - 122

    JO - Molecular Microbiology

    JF - Molecular Microbiology

    SN - 0950-382X

    IS - 1

    ER -