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Summary: Helicase-Dna Polymerase Interaction Is Critical To Initiate Leading-Strand Dna Synthesis | Zhang
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Read the summary and the most important questions on Helicase-DNA polymerase interaction is critical to initiate leading-strand DNA synthesis | Zhang
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1 Abstract
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For what are the interactions between gp4 and gp5 critical?
For initiation of strand displacement synthesis
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2 Introductie
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What is the processivity of gp5/trx alone? And what of gp5/trx interacting with gp4?
800 bp and 17kbp
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What is the processivity of gp4 for DNA unwinding?
60-350 bp
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What is the increase in DNA unwinding speed by gp4 upon interacting with gp5/trx?
From 9.3 - 127 bp/s
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How many basic binding sites does gp5 have?
3, 2 in the TBD (loop A and B) and 1 at the front side
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3 Results
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3.1 Neutralization of a Basic Patch in T7 DNA Polymerase (gp5-Fbpneu) Abolishes Its Ability to Mediate Strand-Displacement DNA Synthesis.
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What is the effect of neutralization of the basic front patch of gp5 on DNA synthesis?
It abolishes the strand-displacement ability of gp5/trx and slows down DNA synthesis by 3 fold
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3.2 The Fbp and the TBD Basic Patch in Gp5 Independently Interact with the C-terminal Tail of Gp4.
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What reduces the interaction between gp5/trx and gp4?
Removal of both the Fbp and TBD of gp5 and removal of the C-termini of gp4
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What is the affinity of gp5/trx for gp4?
Gp5/trx has a Kd of 90nM for gp4
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3.3 Multiple Modes of Interactions Between Gp4 and Gp5/trx in a Replication Complex.
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How many modes of interaction does gp5/trx with gp4 has?
3, 1 via the TBD, 1 via the Fbp and 1 unknown (probably side of gp4)
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3.4 Gp5-Fbpneu∕trx Can Exchange with Gp5/trx at a Replication Fork and Mediate Ongoing Leading-Strand DNA Synthesis.
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What is the effect on interchanging gp5Y526F with gp5FBpneu/trx or gp5/trx?
Both will interchange, but gp5/trx will speed up DNA synthesis in comparison with the other two. Both gp5FBpneu/trx and gp5/trx are discriminating against ddNTPs.
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