caa a22 4500 445818786 CHVBK 20180317145237.0 cr unu---uuuuu 170323e20110301xx s 000 0 eng 10.1007/s00792-010-0352-1 doi (NATIONALLICENCE)springer-10.1007/s00792-010-0352-1 Mutational analysis of conserved aspartic acid residues in the Methanothermobacter thermautotrophicus MCM helicase [Elektronische Daten] [Nozomi Sakakibara, Rajesh Kasiviswanathan, Zvi Kelman] Minichromosome maintenance (MCM) helicases are thought to function as the replicative helicases in archaea and eukarya, unwinding the duplex DNA in the front of the replication fork. The archaeal MCM helicase can be divided into three parts, the N-terminal, catalytic, and C-terminal regions. The N-terminal part of the protein is divided into three domains, A, B, and C, and was shown to be involved in protein multimerization and binding to single- and double-stranded DNA. Two Asp residues found in domain C are conserved among MCM proteins from different archaea. These residues are located in a loop at the interface with domain A. Mutations of these residues in the Methanothermobacter thermautotrophicus MCM protein, Asp202 and Asp203, to Asn result in a significant reduction in the ability of the enzyme to bind DNA and in lower thermal stability. However, the mutant proteins retained helicase and ATPase activities. Further investigation of the DNA binding revealed that the presence of ATP rescues the DNA binding deficiencies by these mutant proteins. Possible roles of these conserved residues in MCM function are discussed. Springer, 2011 Archaea nationallicence DNA replication nationallicence MCM helicase nationallicence Methanothermobacter thermautotrophicus nationallicence CD : Circular dichroism nationallicence dsDNA : Double-stranded DNA nationallicence DSC : Differential scanning calorimetry nationallicence MCM : Minichromosome maintenance nationallicence mt : Methanothermobacter thermautotrophicus nationallicence ssDNA : Single-stranded DNA nationallicence Sakakibara Nozomi Center for Advanced Research in Biotechnology, University of Maryland Biotechnology Institute, 9600 Gudelsky Drive, 20850, Rockville, MD, USA aut Kasiviswanathan Rajesh Center for Advanced Research in Biotechnology, University of Maryland Biotechnology Institute, 9600 Gudelsky Drive, 20850, Rockville, MD, USA aut Kelman Zvi Institute for Bioscience and Biotechnology Research, 9600 Gudelsky Drive, 20850, Rockville, MD, USA aut Extremophiles Springer Japan 15/2(2011-03-01), 245-252 1431-0651 15:2<245 2011 15 792 https://doi.org/10.1007/s00792-010-0352-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00792-010-0352-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sakakibara Nozomi Center for Advanced Research in Biotechnology, University of Maryland Biotechnology Institute, 9600 Gudelsky Drive, 20850, Rockville, MD, USA aut NATIONALLICENCE 700 1- Kasiviswanathan Rajesh Center for Advanced Research in Biotechnology, University of Maryland Biotechnology Institute, 9600 Gudelsky Drive, 20850, Rockville, MD, USA aut NATIONALLICENCE 700 1- Kelman Zvi Institute for Bioscience and Biotechnology Research, 9600 Gudelsky Drive, 20850, Rockville, MD, USA aut NATIONALLICENCE 773 0- Extremophiles Springer Japan 15/2(2011-03-01), 245-252 1431-0651 15:2<245 2011 15 792 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer