Biochemical and biophysical studies on the folding of the core region of the origin of replication of bacteriophage M13.
DNA oligonucleotides with the sequence corresponding to the plus strand origin of replication of the filamentous bacteriophage M13 are studied. Biochemical structure probing and UV melting studies, supplemented with initial NMR experiments, are used to investigate structural features of a 51-nucleotides long synthetic oligonucleotide and two oligonucleotides that are integral parts of this latter molecule. The results demonstrate the feasibility and complementarity of the use of methidiumpropyl.EDTA-Fe(II) and nuclease S1 in the structural analysis of small oligonucleotides. The bacteriophage origin region appears to comprise two hairpins. The first hairpin, which contains a cleavage site for the bacteriophage gene II protein, has a large and probably flexible loop. NMR as well as UV melting studies demonstrate that the second hairpin contains a stable three-membered loop. Both hairpins are present in the 51-mer, which forms a stable tertiary structure.
Department of Biochemistry, University of Leiden, The Netherlands.
A
A
van Belkum
M J
MJ
Blommers
H
H
van den Elst
J H
JH
van Boom
C W
CW
Hilbers
D001435:Bacteriophages / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics / Q000528:radiation effects; D005814:Genes, Viral; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides; D013816:Thermodynamics; D014466:Ultraviolet Rays; D014779:Virus Replication / Q000235:genetics*
D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides