TY - JOUR
T1 - Curved DNA fragments display retarded elution upon anion exchange HPLC
AU - Fack, Fred
AU - Sarantoglou, Vassilis
N1 - Funding Information:
This research was supported by research grants to F.F from the Nato and the Luxemburg Ministry of Education. We thank Dr I. Amiri for using the facilities of the Laboratoire Altergen and the computerized HPLC resources, E.Weiss and C.Kedinger for their SV40 gift, Bernard R£vet for sending us a preprint of his article, Pierre Pfeiffer and Remy Bordonne" for critical reading of the manuscript. We express our gratitude to Eric Westhof for providing us with computer resources and the DNA modeling programs, Guy De Marcillac for precious help for resolving computing problems and Gilbert De Murcia for stimulating discussions and encouragement.
PY - 1991/8/11
Y1 - 1991/8/11
N2 - Restriction fragments containing established curved regions, such as the pBR322 tn3 region, the phage lambda attP region and the SV40 T-antigen and terminus of replication regions, exhibit systematically retarded elution upon anion exchange based HPLC. Using this method, we were able to detect readily other SV40 curved regions, exhibiting also the polyacrylamide gel electrophoresis retardation anomaly, including several RNA polymerase initiation sites. Unlike gel retardation, HPLC retardation exhibited by curved DNA persists at 55δC and is observed for fragments ranging from 150 bp up to 5 kb. The observed preferential attachment of DNA fragments containing curved DNA to the ionic groups of the column suggests a common dipole character of these regions due to the local accumulation of charges resulting apparently from the compression in the minor groove of curved DNA.
AB - Restriction fragments containing established curved regions, such as the pBR322 tn3 region, the phage lambda attP region and the SV40 T-antigen and terminus of replication regions, exhibit systematically retarded elution upon anion exchange based HPLC. Using this method, we were able to detect readily other SV40 curved regions, exhibiting also the polyacrylamide gel electrophoresis retardation anomaly, including several RNA polymerase initiation sites. Unlike gel retardation, HPLC retardation exhibited by curved DNA persists at 55δC and is observed for fragments ranging from 150 bp up to 5 kb. The observed preferential attachment of DNA fragments containing curved DNA to the ionic groups of the column suggests a common dipole character of these regions due to the local accumulation of charges resulting apparently from the compression in the minor groove of curved DNA.
UR - http://www.scopus.com/inward/record.url?scp=0025879860&partnerID=8YFLogxK
U2 - 10.1093/nar/19.15.4181
DO - 10.1093/nar/19.15.4181
M3 - Article
C2 - 1651480
AN - SCOPUS:0025879860
SN - 0305-1048
VL - 19
SP - 4181
EP - 4188
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 15
ER -