Replication origins oriGNAI3 and oriB of the mammalian AMPD2 locus nested in a region of straight DNA flanked by intrinsically bent DNA sites.
The aim of this work was to determine whether intrinsically bent DNA sites are present at, or close to, the mammalian replication origins oriGNAI3 and oriB in the Chinese hamster AMPD2 locus. Using an electrophoretic mobility shift assay and in silico analysis, we located four intrinsically bent DNA sites (b1 to b4) in a fragment that contains the oriGNAI3 and one site (b5) proximal to oriB. The helical parameters show that each bent DNA site is curved in a left-handed superhelical writhe. A 2D projection of 3D fragment trajectories revealed that oriGNAI3 is located in a relatively straight segment flanked by bent sites b1 and b2, which map in previously identified Scaffold/Matrix Attachment Region. Sites b3 and b4 are located approximately 2 kb downstream and force the fragment into a strong closed loop structure. The b5 site is also located in an S/MAR that is found just downstream of oriB.
Departamento de Biologia Celular e Genética, Universidade Estadual de Maringá, UEM, Av. Colombo, 5790, 87020-900, Maringá, Paraná, Brasil.
Valério Américo
VA
Balani
Quirino Alves
QA
de Lima Neto
Karen Izumi
KI
Takeda
Fabrícia
F
Gimenes
Adriana
A
Fiorini
Michelle
M
Debatisse
Maria Aparecida
MA
Fernandez
D000659:AMP Deaminase / Q000235:genetics*; D000818:Animals; D006224:Cricetinae; D004247:DNA / Q000737:chemistry*; D004261:DNA Replication / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D056426:Genetic Loci; D018741:Replication Origin / Q000235:genetics*; D017422:Sequence Analysis, DNA
D004247:DNA; D000659:AMP Deaminase