Identification of ORC1/CDC6-interacting factors in Trypanosoma brucei reveals critical features of origin recognition complex architecture.

DNA replication initiates by formation of a pre-replication complex on sequences termed origins. In eukaryotes, the pre-replication complex is composed of the Origin Recognition Complex (ORC), Cdc6 and the MCM replicative helicase in conjunction with Cdt1. Eukaryotic ORC is considered to be composed...

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Main Authors: Calvin Tiengwe, Lucio Marcello, Helen Farr, Catarina Gadelha, Richard Burchmore, J David Barry, Stephen D Bell, Richard McCulloch
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2012-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0032674&type=printable
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author Calvin Tiengwe
Lucio Marcello
Helen Farr
Catarina Gadelha
Richard Burchmore
J David Barry
Stephen D Bell
Richard McCulloch
author_facet Calvin Tiengwe
Lucio Marcello
Helen Farr
Catarina Gadelha
Richard Burchmore
J David Barry
Stephen D Bell
Richard McCulloch
author_sort Calvin Tiengwe
collection DOAJ
description DNA replication initiates by formation of a pre-replication complex on sequences termed origins. In eukaryotes, the pre-replication complex is composed of the Origin Recognition Complex (ORC), Cdc6 and the MCM replicative helicase in conjunction with Cdt1. Eukaryotic ORC is considered to be composed of six subunits, named Orc1-6, and monomeric Cdc6 is closely related in sequence to Orc1. However, ORC has been little explored in protists, and only a single ORC protein, related to both Orc1 and Cdc6, has been shown to act in DNA replication in Trypanosoma brucei. Here we identify three highly diverged putative T. brucei ORC components that interact with ORC1/CDC6 and contribute to cell division. Two of these factors are so diverged that we cannot determine if they are eukaryotic ORC subunit orthologues, or are parasite-specific replication factors. The other we show to be a highly diverged Orc4 orthologue, demonstrating that this is one of the most widely conserved ORC subunits in protists and revealing it to be a key element of eukaryotic ORC architecture. Additionally, we have examined interactions amongst the T. brucei MCM subunits and show that this has the conventional eukaryotic heterohexameric structure, suggesting that divergence in the T. brucei replication machinery is limited to the earliest steps in origin licensing.
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spelling doaj-art-c3df594cd4bc4893a345aa7b8b3034b72025-08-20T03:09:47ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0173e3267410.1371/journal.pone.0032674Identification of ORC1/CDC6-interacting factors in Trypanosoma brucei reveals critical features of origin recognition complex architecture.Calvin TiengweLucio MarcelloHelen FarrCatarina GadelhaRichard BurchmoreJ David BarryStephen D BellRichard McCullochDNA replication initiates by formation of a pre-replication complex on sequences termed origins. In eukaryotes, the pre-replication complex is composed of the Origin Recognition Complex (ORC), Cdc6 and the MCM replicative helicase in conjunction with Cdt1. Eukaryotic ORC is considered to be composed of six subunits, named Orc1-6, and monomeric Cdc6 is closely related in sequence to Orc1. However, ORC has been little explored in protists, and only a single ORC protein, related to both Orc1 and Cdc6, has been shown to act in DNA replication in Trypanosoma brucei. Here we identify three highly diverged putative T. brucei ORC components that interact with ORC1/CDC6 and contribute to cell division. Two of these factors are so diverged that we cannot determine if they are eukaryotic ORC subunit orthologues, or are parasite-specific replication factors. The other we show to be a highly diverged Orc4 orthologue, demonstrating that this is one of the most widely conserved ORC subunits in protists and revealing it to be a key element of eukaryotic ORC architecture. Additionally, we have examined interactions amongst the T. brucei MCM subunits and show that this has the conventional eukaryotic heterohexameric structure, suggesting that divergence in the T. brucei replication machinery is limited to the earliest steps in origin licensing.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0032674&type=printable
spellingShingle Calvin Tiengwe
Lucio Marcello
Helen Farr
Catarina Gadelha
Richard Burchmore
J David Barry
Stephen D Bell
Richard McCulloch
Identification of ORC1/CDC6-interacting factors in Trypanosoma brucei reveals critical features of origin recognition complex architecture.
PLoS ONE
title Identification of ORC1/CDC6-interacting factors in Trypanosoma brucei reveals critical features of origin recognition complex architecture.
title_full Identification of ORC1/CDC6-interacting factors in Trypanosoma brucei reveals critical features of origin recognition complex architecture.
title_fullStr Identification of ORC1/CDC6-interacting factors in Trypanosoma brucei reveals critical features of origin recognition complex architecture.
title_full_unstemmed Identification of ORC1/CDC6-interacting factors in Trypanosoma brucei reveals critical features of origin recognition complex architecture.
title_short Identification of ORC1/CDC6-interacting factors in Trypanosoma brucei reveals critical features of origin recognition complex architecture.
title_sort identification of orc1 cdc6 interacting factors in trypanosoma brucei reveals critical features of origin recognition complex architecture
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0032674&type=printable
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