Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1099/jgv.0.001253
Titel (primär) Secondary structure of DNA released from purified capsids of human parvovirus B19 under moderate denaturing conditions
Autor Matz, B.; Kupfer, B.; Kallies, R.; Külshammer, M.; Flötenmeyer, M.; Kreil, T.R.; Eis-Hübinger, A.M.
Quelle Journal of General Virology
Erscheinungsjahr 2019
Department UMB
Band/Volume 100
Heft 5
Seite von 812
Seite bis 827
Sprache englisch
Keywords B19V; secondary genome structure; B19V DNA; Parvovirus B19
Abstract Parvovirus B19 (B19V) possesses a linear single-stranded DNA genome of either positive or negative polarity. Due to intramolecular sequence homologies, either strand may theoretically be folded in several alternative ways. Viral DNA, when extracted from virions by several procedures, presents as linear single-stranded and/or linear double-stranded molecules, except when one particular commercial kit is used. This protocol yields DNA with an aberrant electrophoretic mobility in addition to linear double-stranded molecules, but never any single-stranded molecules. This peculiar kind of DNA was found in all plasma or serum samples tested and so we decided to analyse its secondary structure. In line with our results for one- and two-dimensional electrophoresis, mobility shift assays, DNA preparation by an in-house extraction method with moderate denaturing conditions, density gradient ultracentrifugation, DNA digestion experiments and competition hybridization assays, we conclude that (i) the unique internal portions of this distinctive single-stranded molecules are folded into tight tangles and (ii) the two terminal redundant regions are associated with each other, yielding non-covalently closed pseudo-circular molecules stabilized by a short (18 nucleotides) intramolecular stem, whereas the extreme 3′- and 5′-ends are folded back on themselves, forming a structure resembling a twin hairpin. The question arises as to whether this fairly unstable structure represents the encapsidated genome structure. The answer to this question remains quite relevant in terms of comprehending the initiation and end of B19V genome replication.
dauerhafte UFZ-Verlinkung
Matz, B., Kupfer, B., Kallies, R., Külshammer, M., Flötenmeyer, M., Kreil, T.R., Eis-Hübinger, A.M. (2019):
Secondary structure of DNA released from purified capsids of human parvovirus B19 under moderate denaturing conditions
J. Gen. Virol. 100 (5), 812 - 827 10.1099/jgv.0.001253