DNA binding properties of human Cdc45 suggest a function as molecular wedge for DNA unwinding
Please always quote using this URN: urn:nbn:de:bvb:20-opus-117538
- The cell division cycle protein 45 (Cdc45) represents an essential replication factor that, together with the Mcm2-7 complex and the four subunits of GINS, forms the replicative DNA helicase in eukaryotes. Recombinant human Cdc45 (hCdc45) was structurally characterized and its DNA-binding properties were determined. Synchrotron radiation circular dichroism spectroscopy, dynamic light scattering, small-angle X-ray scattering and atomic force microscopy revealed that hCdc45 exists as an alpha-helical monomer and possesses a structure similar toThe cell division cycle protein 45 (Cdc45) represents an essential replication factor that, together with the Mcm2-7 complex and the four subunits of GINS, forms the replicative DNA helicase in eukaryotes. Recombinant human Cdc45 (hCdc45) was structurally characterized and its DNA-binding properties were determined. Synchrotron radiation circular dichroism spectroscopy, dynamic light scattering, small-angle X-ray scattering and atomic force microscopy revealed that hCdc45 exists as an alpha-helical monomer and possesses a structure similar to its bacterial homolog RecJ. hCdc45 bound long (113-mer or 80-mer) single-stranded DNA fragments with a higher affinity than shorter ones (34-mer). hCdc45 displayed a preference for 3' protruding strands and bound tightly to single-strand/double-strand DNA junctions, such as those presented by Y-shaped DNA, bubbles and displacement loops, all of which appear transiently during the initiation of DNA replication. Collectively, our findings suggest that hCdc45 not only binds to but also slides on DNA with a 3'-5' polarity and, thereby acts as a molecular 'wedge' to initiate DNA strand displacement.…
Author: | Anna Szambowska, Ingrid Tessmer, Petri Kursula, Christian Usskilat, Potr Prus, Helmut Pospiech, Frank Grosse |
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URN: | urn:nbn:de:bvb:20-opus-117538 |
Document Type: | Journal article |
Faculties: | Fakultät für Biologie / Julius-von-Sachs-Institut für Biowissenschaften |
Language: | English |
Parent Title (English): | Nucleic Acids Research |
ISSN: | 1362-4962 |
Year of Completion: | 2014 |
Volume: | 42 |
Issue: | 4 |
Pagenumber: | 2308-2319 |
Source: | Nucleic Acids Research, 2014, Vol. 42, No. 4, 2308-2319. doi:10.1093/nar/gkt1217 |
DOI: | https://doi.org/10.1093/nar/gkt1217 |
Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/24293646 |
Dewey Decimal Classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
Tag: | circular dichroism spectra; cyclin-dependent kinases; escherichia coli; protein secondary structure; ray solution scattering; recj exonuclease; s-phase; saccharmyces cerevisiae; single-stranded-DNA; small-angle scattering |
Release Date: | 2015/08/24 |
Licence (German): | CC BY: Creative-Commons-Lizenz: Namensnennung |