Human origins of DNA replication selected from a library of nascent DNA.

Abstract:

:The identification of metazoan origins of DNA replication has so far been hampered by the lack of a suitable genetic screening and by the cumbersomeness of the currently available mapping procedures. Here we describe the construction of a library of nascent DNA, representative of all cellular origin sequences, and its utilization as a screening probe for origin identification in large genomic regions. The procedure developed was successfully applied to the human 5q31.1 locus, encoding for the IL-3 and GM-CSF genes. Two novel origins were identified and subsequently characterized by competitive PCR mapping, located approximately 3.5 kb downstream of the GM-CSF gene. The two origins (GM-CSF Ori1 and Ori2) were shown to interact with different members of the DNA prereplication complex. This observation reinforces the universal paradigm that initiation of DNA replication takes place at, or in close proximity to, the binding sites of the trans-acting initiator proteins.

journal_name

Mol Cell

journal_title

Molecular cell

authors

Todorovic V,Giadrossi S,Pelizon C,Mendoza-Maldonado R,Masai H,Giacca M

doi

10.1016/j.molcel.2005.07.005

subject

Has Abstract

pub_date

2005-08-19 00:00:00

pages

567-75

issue

4

eissn

1097-2765

issn

1097-4164

pii

S1097-2765(05)01443-7

journal_volume

19

pub_type

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