Directional motion of foreign plasmid DNA to nuclear HP1 foci.
Movement of labelled plasmid DNA relative to heterochromatin foci in nuclei, visualized with HP1-GFP, was studied using live-cell imaging and object tracking. In addition to Brownian motion of plasmid DNA we found a pronounced, non-random movement of plasmid DNA towards the nearest HP1 focus, while time-lapse microscopy showed that HP1 foci are relatively immobile and positionally stable. The movement of plasmid DNA was much faster than that of the HP1 foci. Contact of transgene DNA with an HP1 focus usually resulted in cessation of the directional motion. Moreover, the motion of plasmid DNA inside the heterochromatin compartment was more restricted (limited to 0.25 microm) than when the plasmid DNA was outside heterochromatin (R = 0.7 microm). Three days after transfection most of the foreign labelled DNA colocalized with centromeric heterochromatin.
Ondrej
Vladan
V
Laboratory of Molecular Cytology and Cytometry, Institute of Biophysics AS CR, Kralovopolska 135, Brno, 612 65, Czech Republic. ondrej@ibp.cz
Kozubek
Stanislav
S
Lukásová
Emílie
E
Falk
Martin
M
Matula
Pavel
P
Matula
Petr
P
Kozubek
Michal
M
D001692:Biological Transport / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000502:physiology; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000502:physiology*; D004247:DNA / Q000235:genetics / Q000502:physiology*; D006570:Heterochromatin; D006801:Humans; D008853:Microscopy; D010957:Plasmids / Q000235:genetics / Q000502:physiology*; D014162:Transfection
D002868:Chromosomal Proteins, Non-Histone; D006570:Heterochromatin; D000090266:Chromobox Protein Homolog 5; D004247:DNA