Identification of eukaryotic promoter regulatory elements using nonhomologous random recombination.

Nucleic Acids Res
Authors
Keywords
Abstract

Understanding the regulatory logic of a eukaryotic promoter requires the elucidation of the regulatory elements within that promoter. Current experimental or computational methods to discover regulatory motifs within a promoter can be labor intensive and may miss redundant, unprecedented or weakly activating elements. We have developed an unbiased combinatorial approach to rapidly identify new upstream activating sequences (UASs) in a promoter. This approach couples nonhomologous random recombination with an in vivo screen to efficiently identify UASs and does not rely on preconceived hypotheses about promoter regulation or on similarity to known activating sequences. We validated this method using the unfolded protein response (UPR) in yeast and were able to identify both known and potentially novel UASs involved in the UPR. One of the new UASs discovered using this approach implicates Crz1 as a possible activator of Hac1, a transcription factor involved in the UPR. This method has several advantages over existing methods for UAS discovery including its speed, potential generality, sensitivity and lack of false positives and negatives.

Year of Publication
2007
Journal
Nucleic Acids Res
Volume
35
Issue
17
Pages
5851-60
Date Published
2007
ISSN
1362-4962
DOI
10.1093/nar/gkm634
PubMed ID
17720707
PubMed Central ID
PMC2034452
Links
Grant list
R01 GM065400 / GM / NIGMS NIH HHS / United States