Publications
58 results found
Riera A, Tognetti S, Speck C, 2014, Helicase loading: How to build a MCM2-7 double-hexamer, Seminars in Cell and Developmental Biology, Vol: 30, Pages: 104-109, ISSN: 1084-9521
A central step in eukaryotic initiation of DNA replication is the loading of the helicase at replication origins, misregulation of this reaction leads to DNA damage and genome instability. Here we discuss how the helicase becomes recruited to origins and loaded into a double-hexamer around double-stranded DNA. We specifically describe the individual steps in complex assembly and explain how this process is regulated to maintain genome stability. Structural analysis of the helicase loader and the helicase has provided key insights into the process of double-hexamer formation. A structural comparison of the bacterial and eukaryotic system suggests a mechanism of helicase loading. © 2014 Elsevier Ltd.
Riera A, Li H, Speck C, 2013, The MCM2-7 helicase trapped in complex with its DNA loader, Cell Cycle, Vol: 12, Pages: 2917-2918, ISSN: 1538-4101
Riera A, Fernandez-Cid A, Speck C, 2013, The ORC/Cdc6/MCM2-7 complex, a new power player for regulated helicase loading, CELL CYCLE, Vol: 12, Pages: 2155-2156, ISSN: 1538-4101
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- Citations: 5
Sun J, Evrin C, Samel AS, et al., 2013, Cryo-EM structure of a helicase loading intermediate containing ORC-Cdc6-Cdt1-MCM2-7 bound to DNA, Nature Structure Molecular Biology
Fernandez-Cid A, Riera A, Tognetti S, et al., 2013, An ORC/Cdc6/MCM2-7 Complex Is Formed in a Multistep Reaction to Serve as a Platform for MCM Double-Hexamer Assembly, MOLECULAR CELL, Vol: 50, Pages: 577-588, ISSN: 1097-2765
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- Citations: 99
Evrin C, Fernández-Cid A, Zech J, et al., 2013, In the absence of ATPase activity, pre-RC formation is blocked prior to MCM2–7 hexamer dimerization, Nucleic Acids Research
Sun J, Kawakami H, Zech J, et al., 2012, Cdc6-Induced Conformational Changes in ORC Bound to Origin DNA Revealed by Cryo-Electron Microscopy, STRUCTURE, Vol: 20, Pages: 534-544, ISSN: 0969-2126
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- Citations: 52
Chabes A, Speck C, Johansson E, 2011, A top-down view on DNA replication and recombination from 9,000 feet above sea level., Genome Biology: biology for the post-genomic era, Vol: 12, Pages: 1-2, ISSN: 1474-7596
A report of the Keystone Symposium 'DNA Replication and Recombination' held in Keystone, USA, 27 February to 4 March 2011.
Evrin C, Clarke P, Zech J, et al., 2009, A double-hexameric MCM2-7 complex is loaded onto origin DNA during licensing of eukaryotic DNA replication, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol: 106, Pages: 20240-20245, ISSN: 0027-8424
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- Citations: 373
Chen Z, Speck C, Wendel P, et al., 2008, The architecture of the DNA replication origin recognition complex in Saccharomyces cerevisiae., Proceedings of the National Academy of Sciences of USA
Speck C, Stillman B, 2007, Initiation of DNA replication - Assembly of a molecular machine depends on origin DNA, 32nd Congress of the Federation-of-European-Biochemical-Societies (FEBS), Publisher: BLACKWELL PUBLISHING, Pages: 72-72, ISSN: 1742-464X
Speck C, Stillman B, 2007, Cdc6 ATPase activity regulates ORC x Cdc6 stability and the selection of specific DNA sequences as origins of DNA replication., J Biol Chem, Vol: 282, Pages: 11705-11714, ISSN: 0021-9258
DNA replication, as with all macromolecular synthesis steps, is controlled in part at the level of initiation. Although the origin recognition complex (ORC) binds to origins of DNA replication, it does not solely determine their location. To initiate DNA replication ORC requires Cdc6 to target initiation to specific DNA sequences in chromosomes and with Cdt1 loads the ring-shaped mini-chromosome maintenance (MCM) 2-7 DNA helicase component onto DNA. ORC and Cdc6 combine to form a ring-shaped complex that contains six AAA+ subunits. ORC and Cdc6 ATPase mutants are defective in MCM loading, and ORC ATPase mutants have reduced activity in ORC x Cdc6 x DNA complex formation. Here we analyzed the role of the Cdc6 ATPase on ORC x Cdc6 complex stability in the presence or absence of specific DNA sequences. Cdc6 ATPase is activated by ORC, regulates ORC x Cdc6 complex stability, and is suppressed by origin DNA. Mutations in the conserved origin A element, and to a lesser extent mutations in the B1 and B2 elements, induce Cdc6 ATPase activity and prevent stable ORC x Cdc6 formation. By analyzing ORC x Cdc6 complex stability on various DNAs, we demonstrated that specific DNA sequences control the rate of Cdc6 ATPase, which in turn controls the rate of Cdc6 dissociation from the ORC x Cdc6 x DNA complex. We propose a mechanism explaining how Cdc6 ATPase activity promotes origin DNA sequence specificity; on DNA that lacks origin activity, Cdc6 ATPase promotes dissociation of Cdc6, whereas origin DNA down-regulates Cdc6 ATPase resulting in a stable ORC x Cdc6 x DNA complex, which can then promote MCM loading. This model has relevance for origin specificity in higher eukaryotes.
Majka J, Speck C, 2007, Analysis of protein-DNA interactions using surface plasmon resonance., Adv Biochem Eng Biotechnol, Vol: 104, Pages: 13-36, ISSN: 0724-6145
Protein-DNA interactions are required for access and protection of the genetic information within the cell. Historically these interactions have been studied using genetic, biochemical, and structural methods resulting in qualitative or semiquantitative interaction data. In the future the focus will be on high quality quantitative data to model a huge number of interactions forming a specific network in system biology approaches. Toward this aim, BIAcore introduced in 1990 the first commercial machine that uses surface plasmon resonance (SPR) to study the real-time kinetics of biomolecular interactions. Since then systems have been developed to allow for robust analysis of a multitude of protein-DNA interactions. Here we provide a detailed guide for protein-DNA interaction analysis using the BIAcore, starting with a description of the SPR technology, giving recommendations on preliminary studies, and finishing with extensive information on quantitative and qualitative data analysis. One focus is on cooperative protein-DNA interactions, where proteins interact with each other to modulate their binding specificity or affinity. The BIAcore has been used for the last 14 years to study protein-DNA interactions; our literature review focuses on some high quality studies describing a wide range of experimental uses, covering simple 1 : 1 interactions, analysis of complicated multiprotein-DNA interaction systems, and analytical uses.
Majka J, Speck C, 2006, Analysis of protein-DNA interactions using surface plasmon resonance, Advances in Biochemical Engineering/Biotechnology, Vol: 104, Pages: 13-36, ISSN: 0724-6145
Protein-DNA interactions are required for access and protection of the genetic information within the cell. Historically these interactions have been studied using genetic, biochemical, and structural methods resulting in qualitative or semiquantitative interaction data. In the future the focus will be on high quality quantitative data to model a huge number of interactions forming a specific network in system biology approaches. Toward this aim, BIAcore introduced in 1990 the first commercial machine that uses surface plasmon resonance (SPR) to study the real-time kinetics of biomolecular interactions. Since then systems have been developed to allow for robust analysis of a multitude of protein-DNA interactions. Here we provide a detailed guide for protein-DNA interaction analysis using the BIAcore, starting with a description of the SPR technology, giving recommendations on preliminary studies, and finishing with extensive information on quantitative and qualitative data analysis. One focus is on cooperative protein-DNA interactions, where proteins interact with each other to modulate their binding specificity or affinity. The BIAcore has been used for the last 14 years to study protein-DNA interactions; our literature review focuses on some high quality studies describing a wide range of experimental uses, covering simple 1:1 interactions, analysis of complicated multiprotein-DNA interaction systems, and analytical uses. © Springer-Verlag Berlin Heidelberg 2006.
Speck C, Chen Z, Li H, et al., 2005, ATPase-dependent cooperative binding of ORC and Cdc6 to origin DNA., Nat Struct Mol Biol, Vol: 12, Pages: 965-971, ISSN: 1545-9993
Binding of Cdc6 to the origin recognition complex (ORC) is a key step in the assembly of a pre-replication complex (pre-RC) at origins of DNA replication. ORC recognizes specific origin DNA sequences in an ATP-dependent manner. Here we demonstrate cooperative binding of Saccharomyces cerevisiae Cdc6 to ORC on DNA in an ATP-dependent manner, which induces a change in the pattern of origin binding that requires the Orc1 ATPase. The reaction is blocked by specific origin mutations that do not interfere with the interaction between ORC and DNA. Single-particle reconstruction of electron microscopic images shows that the ORC-Cdc6 complex forms a ring-shaped structure with dimensions similar to those of the ring-shaped MCM helicase. The ORC-Cdc6 structure is predicted to contain six AAA+ subunits, analogous to other ATP-dependent protein machines. We suggest that Cdc6 and origin DNA activate a molecular switch in ORC that contributes to pre-RC assembly.
de la Hoz AB, Pratto F, Misselwitz R, et al., 2004, Recognition of DNA by ω protein from the broad-host range <i>Streptococcus pyogenes</i> plasmid pSM19035:: analysis of binding to operator DNA with one to four heptad repeats, NUCLEIC ACIDS RESEARCH, Vol: 32, Pages: 3136-3147, ISSN: 0305-1048
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- Citations: 40
Berenstein D, Olesen K, Speck C, et al., 2002, Genetic organization of the <i>Vibrio harveyi dnaA</i> gene region and analysis of the function of the <i>V-harveyi</i> DnaA protein in <i>Escherichia coli</i>, JOURNAL OF BACTERIOLOGY, Vol: 184, Pages: 2533-+, ISSN: 0021-9193
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- Citations: 11
Pieper U, Groll DH, Wünsch S, et al., 2002, The GTP-dependent restriction enzyme McrBC from <i>Escherichia coli</i> forms high-molecular mass complexes with DNA and produces a cleavage pattern with a characteristic 10-base pair repeat, BIOCHEMISTRY, Vol: 41, Pages: 5245-5254, ISSN: 0006-2960
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- Citations: 23
Speck C, Messer W, 2001, Mechanism of origin unwinding: sequential binding of DnaA to double- and single-stranded DNA, EMBO JOURNAL, Vol: 20, Pages: 1469-1476, ISSN: 0261-4189
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- Citations: 166
Messer W, Blaesing F, Jakimowicz D, et al., 2001, Bacterial replication initiator DnaA. Rules for DnaA binding and roles of DnaA in origin unwinding and helicase loading, BIOCHIMIE, Vol: 83, Pages: 5-12, ISSN: 0300-9084
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- Citations: 83
Schaper S, Nardmann J, Lüder G, et al., 2000, Identification of the chromosomal replication origin from <i>Thermos thermophilus</i> and its interaction with the replication initiator DnaA, JOURNAL OF MOLECULAR BIOLOGY, Vol: 299, Pages: 655-665, ISSN: 0022-2836
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- Citations: 28
Skarstad K, Lueder G, Lurz R, et al., 2000, The <i>Escherichia coli</i> SeqA protein binds specifically and co-operatively to two sites in hemimethylated and fully methylated <i>oriC</i>, MOLECULAR MICROBIOLOGY, Vol: 36, Pages: 1319-1326, ISSN: 0950-382X
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- Citations: 55
Kohler M, Speck C, Christiansen M, et al., 1999, Evidence for distinct substrate specificities of importin alpha family members in nuclear protein import, MOLECULAR AND CELLULAR BIOLOGY, Vol: 19, Pages: 7782-7791, ISSN: 0270-7306
Speck C, Weigel C, Messer W, 1999, ATP- and ADP-DnaA protein, a molecular switch in gene regulation, EMBO JOURNAL, Vol: 18, Pages: 6169-6176, ISSN: 0261-4189
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- Citations: 153
Messer W, Blaesing F, Majka J, et al., 1999, Functional domains of DnaA proteins, Bacterial Cell Cycle Meeting, Publisher: EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER, Pages: 819-825, ISSN: 0300-9084
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- Citations: 77
Szalewska-Palasz A, Weigel C, Speck C, et al., 1998, Interaction of the <i>Escherichia coli</i> DnaA protein with bacteriophage λ DNA, MOLECULAR AND GENERAL GENETICS, Vol: 259, Pages: 679-688, ISSN: 0026-8925
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- Citations: 24
Jakimowicz D, Majka J, Messer W, et al., 1998, Structural elements of the <i>Streptomyces oriC</i> region and their interactions with the DnaA protein, MICROBIOLOGY-SGM, Vol: 144, Pages: 1281-1290, ISSN: 1350-0872
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- Citations: 60
Speck C, Weigel C, Messer W, 1997, From footprint to toeprint: A close-up of the DnaA box, the binding site for the bacterial initiator protein DnaA, NUCLEIC ACIDS RESEARCH, Vol: 25, Pages: 3242-3247, ISSN: 0305-1048
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- Citations: 22
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