Hsorc4-dependent DNA remodeling of the ori-beta dhfr replicator
Abstract
Replication of DNA in multicellular organisms initiates from origin of replication (ori) sequences, which significantly differ in length and complexity. One of the best characterized is hamster dihydrofolate reductase (DHFR), which contains the ori-beta sequence with several functionally relevant domains, such as an AT-rich region, dinucleotide repeat element (DNR), sequence-induced bend DNA (BEND) and a RIP60 protein-binding site (RIP60). Prior to initiation, ori sequences are recognized by origin recognition complex (ORC), which is a hetero hexamer complex that serves as the landing pad for proteins of the pre-replication complex. The function of each ORC subunit is still unclear. In this study, we analyze the function of subunit 4 of the human ORC complex (HsOrc4) in interaction with a plasmid bearing the ori-beta DHFR sequence. We show that the topologically closed DHFR ori-beta replicator contains a bubble-like structure within its AT-rich region and that it is reversibly modified... in the interaction with HsOrc4. The non-canonical structure of the AT-rich region in the topologically closed ori sequence is recognized and changed by HsOrc4 using the energy of supercoiled DNA. These findings could help to further elucidate DNA replication and its possible association with human genetic diseases.
Keywords:
Topoisomerase I / Supercoiled plasmid / Replicator / ori-beta DHFR / ORC complex / Mung bean nuclease / HsOrc4 / DNA topologySource:
Cellular & Molecular Biology Letters, 2015, 20, 4, 549-561Publisher:
- Walter De Gruyter Gmbh, Berlin
Funding / projects:
- Complex diseases as a model system for phenotype modulation- structural and functional analysis of molecular biomarkers (RS-MESTD-Basic Research (BR or ON)-173008)
- International Centre for Genetic Engineering and Biotechnology in Italy [CRP/YUG08-01]
DOI: 10.1515/cmble-2015-0032
ISSN: 1425-8153
PubMed: 26124052
WoS: 000363055000002
Scopus: 2-s2.0-84946240219
Collections
Institution/Community
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Tomić, Branko AU - Kušić-Tišma, Jelena PY - 2015 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/816 AB - Replication of DNA in multicellular organisms initiates from origin of replication (ori) sequences, which significantly differ in length and complexity. One of the best characterized is hamster dihydrofolate reductase (DHFR), which contains the ori-beta sequence with several functionally relevant domains, such as an AT-rich region, dinucleotide repeat element (DNR), sequence-induced bend DNA (BEND) and a RIP60 protein-binding site (RIP60). Prior to initiation, ori sequences are recognized by origin recognition complex (ORC), which is a hetero hexamer complex that serves as the landing pad for proteins of the pre-replication complex. The function of each ORC subunit is still unclear. In this study, we analyze the function of subunit 4 of the human ORC complex (HsOrc4) in interaction with a plasmid bearing the ori-beta DHFR sequence. We show that the topologically closed DHFR ori-beta replicator contains a bubble-like structure within its AT-rich region and that it is reversibly modified in the interaction with HsOrc4. The non-canonical structure of the AT-rich region in the topologically closed ori sequence is recognized and changed by HsOrc4 using the energy of supercoiled DNA. These findings could help to further elucidate DNA replication and its possible association with human genetic diseases. PB - Walter De Gruyter Gmbh, Berlin T2 - Cellular & Molecular Biology Letters T1 - Hsorc4-dependent DNA remodeling of the ori-beta dhfr replicator EP - 561 IS - 4 SP - 549 VL - 20 DO - 10.1515/cmble-2015-0032 ER -
@article{ author = "Tomić, Branko and Kušić-Tišma, Jelena", year = "2015", abstract = "Replication of DNA in multicellular organisms initiates from origin of replication (ori) sequences, which significantly differ in length and complexity. One of the best characterized is hamster dihydrofolate reductase (DHFR), which contains the ori-beta sequence with several functionally relevant domains, such as an AT-rich region, dinucleotide repeat element (DNR), sequence-induced bend DNA (BEND) and a RIP60 protein-binding site (RIP60). Prior to initiation, ori sequences are recognized by origin recognition complex (ORC), which is a hetero hexamer complex that serves as the landing pad for proteins of the pre-replication complex. The function of each ORC subunit is still unclear. In this study, we analyze the function of subunit 4 of the human ORC complex (HsOrc4) in interaction with a plasmid bearing the ori-beta DHFR sequence. We show that the topologically closed DHFR ori-beta replicator contains a bubble-like structure within its AT-rich region and that it is reversibly modified in the interaction with HsOrc4. The non-canonical structure of the AT-rich region in the topologically closed ori sequence is recognized and changed by HsOrc4 using the energy of supercoiled DNA. These findings could help to further elucidate DNA replication and its possible association with human genetic diseases.", publisher = "Walter De Gruyter Gmbh, Berlin", journal = "Cellular & Molecular Biology Letters", title = "Hsorc4-dependent DNA remodeling of the ori-beta dhfr replicator", pages = "561-549", number = "4", volume = "20", doi = "10.1515/cmble-2015-0032" }
Tomić, B.,& Kušić-Tišma, J.. (2015). Hsorc4-dependent DNA remodeling of the ori-beta dhfr replicator. in Cellular & Molecular Biology Letters Walter De Gruyter Gmbh, Berlin., 20(4), 549-561. https://doi.org/10.1515/cmble-2015-0032
Tomić B, Kušić-Tišma J. Hsorc4-dependent DNA remodeling of the ori-beta dhfr replicator. in Cellular & Molecular Biology Letters. 2015;20(4):549-561. doi:10.1515/cmble-2015-0032 .
Tomić, Branko, Kušić-Tišma, Jelena, "Hsorc4-dependent DNA remodeling of the ori-beta dhfr replicator" in Cellular & Molecular Biology Letters, 20, no. 4 (2015):549-561, https://doi.org/10.1515/cmble-2015-0032 . .