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 Celljournal_title
Molecular cellauthors
Todorovic V,Giadrossi S,Pelizon C,Mendoza-Maldonado R,Masai H,Giacca Mdoi
10.1016/j.molcel.2005.07.005subject
Has Abstractpub_date
2005-08-19 00:00:00pages
567-75issue
4eissn
1097-2765issn
1097-4164pii
S1097-2765(05)01443-7journal_volume
19pub_type
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