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Wild type and DDK phosphorylated MCM2 demonstrate unique binding specificities for RLF2 and CAC2 respectively.

Title: Wild type and DDK phosphorylated MCM2 demonstrate unique binding specificities for RLF2 and CAC2 respectively.
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Name(s): Jones, John Matthew, author
Type of Resource: text
Genre: Text
Date Issued: 2016-12-09
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: Minichromosome maintenance complex 2-7 (MCM2-7) is a fundamental component of the replicative CMG helicase (CDC45-MCM-GINS) characteristic of active eukaryotic replication forks. MCM2-7 is composed of 6 subunits, some of which are capable of receiving Dbf4-Cdc7 kinase (DDK) phosphorylation. Specifically, one of the six MCM2-7 subunits, MCM2, is associated with CMG complex activation upon MCM2 phosphorylation from DDK. Dr. Daniel Kaplan of Florida State University has developed an MCM2 phosphomutant (MCM2 11e) shown to mimic DDK phosphorylated MCM2, and therefore MCM2 that is present in active CMG helicase complexes [19]. Recent research of MCM2 has suggested accessory activities of the MCM2 protein aside from participation in the MCM2-7 ring of the CMG helicase complex. One particular function is histone chaperoning of H3-H4 dimers, a characteristic shared by chromatin assembly factor 1 (CAF-1). CAF-1 is known to deposit H3-H4 dimers at the replication fork, providing enough suspicion to investigate MCM2-CAF-1 interactions. In this Honors Thesis, GST pull-down assays were conducted to test MCM2 wild type (wt) and 11e binding efficiencies for 2 of 3 CAF-1 subunits, RLF2 and CAC2. Results indicate significant MCM2 wt affinity for RLF2, affinity not reciprocated by MCM2 11e. However, neither MCM2 wt nor 11e exhibit binding efficiencies greater than solely GST tag controls do for CAC2. The preferential binding of MCM2 wt to RLF2 and the shared repulsion of CAC2 by MCM2 wt and 11e identify MCM2 binding specificity to CAF-1 subunits not previously identified.
Identifier: FSU_libsubv1_scholarship_submission_1481561146 (IID)
Keywords: MCM2, CAF-1, Minichromosome, Maintanance, Complex, 11e, S164D, S170D, DNA, H3-H4, John Matthew Jones, John, Matthew, Jones, chaperone, chaperoning, MCM2 11e, MCM2-7, replication, replication fork, fork, helicase, ASF1, CAC2, RLF2, Daniel, Kaplan,
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_libsubv1_scholarship_submission_1481561146
Owner Institution: FSU

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Jones, J. M. (2016). Wild type and DDK phosphorylated MCM2 demonstrate unique binding specificities for RLF2 and CAC2 respectively. Retrieved from http://purl.flvc.org/fsu/fd/FSU_libsubv1_scholarship_submission_1481561146