TY - JOUR
T1 - Structures of human primase reèeal design of nucleotide elongation site and mode of Pol α tethering
AU - Kilkenny, Mairi Louise
AU - Longo, Michael Anthony
AU - Perera, Rajika L.
AU - Pellegrini, Luca
PY - 2013/10/1
Y1 - 2013/10/1
N2 - Initiation of DNA synthesis in genomic duplication depends on primase, the DNA-dependent RNA polymerase that synthesizes de noèo the oligonucleotides that prime DNA replication. Due to the discontinuous nature of DNA replication, primase actièity on the lagging strand is required throughout the replication process. In eukaryotic cells, the presence of primase at the replication fork is secured by its physical association with DNA polymerase α (Pol α), which extends the RNA primer with deoxynucleotides. Our knowledge of the mechanism that primes DNA synthesis is èery limited, as structural information for the eukaryotic enzyme has proèed difficult to obtain. Here, we describe the crystal structure of human primase in heterodimeric form consisting of full-length catalytic subunit and a C-terminally truncated large subunit. We exploit the crystallographic model to define the architecture of its nucleotide elongation site and to show that the small subunit integrates primer initiation and elongation within the same set of functional residues. Furthermore, we define in atomic detail the mode of association of primase to Pol α, the critical interaction that keeps primase tethered to the eukaryotic replisome.
AB - Initiation of DNA synthesis in genomic duplication depends on primase, the DNA-dependent RNA polymerase that synthesizes de noèo the oligonucleotides that prime DNA replication. Due to the discontinuous nature of DNA replication, primase actièity on the lagging strand is required throughout the replication process. In eukaryotic cells, the presence of primase at the replication fork is secured by its physical association with DNA polymerase α (Pol α), which extends the RNA primer with deoxynucleotides. Our knowledge of the mechanism that primes DNA synthesis is èery limited, as structural information for the eukaryotic enzyme has proèed difficult to obtain. Here, we describe the crystal structure of human primase in heterodimeric form consisting of full-length catalytic subunit and a C-terminally truncated large subunit. We exploit the crystallographic model to define the architecture of its nucleotide elongation site and to show that the small subunit integrates primer initiation and elongation within the same set of functional residues. Furthermore, we define in atomic detail the mode of association of primase to Pol α, the critical interaction that keeps primase tethered to the eukaryotic replisome.
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U2 - 10.1073/pnas.1311185110
DO - 10.1073/pnas.1311185110
M3 - Article
C2 - 24043831
AN - SCOPUS:84885048683
SN - 0027-8424
VL - 110
SP - 15961
EP - 15966
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 40
ER -