Acid nuclear extracts as mediators of gene transfer and expression.
In an attempt to demonstrate transfection-active DNA packaging proteins in the cell nucleus, we prepared acid nuclear extracts with perchloric acid and subsequent protein fractions by stepwise acetone precipitation. The original extract and these fractions containing different compositions of nuclear proteins were used as DNA packaging agents. After the formation of complexes between these protein fractions and reporter genes, the addition of these complexes to the cells resulted in high transfection rates. Gel electrophoresis shows that the most active fractions contain histone H1 and HMG17. HMG1 exhibits a smaller activity. This result was confirmed by positive transfection experiments with commercial histone H1. Our results show that the transfection activity of acid nuclear protein fractions and histone H1 is dependent on the presence of calcium.
Böttger
M
M
Nephrology, Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany. mboettg@mdc-berlin.de
Zaitsev
S V
SV
Otto
A
A
Haberland
A
A
Vorob'ev
V I
VI
D016475:3T3 Cells; D000818:Animals; D002118:Calcium; D002417:Cattle; D002457:Cell Extracts; D002467:Cell Nucleus / Q000737:chemistry*; D003470:Culture Media; D004705:Endocytosis; D006609:High Mobility Group Proteins; D006657:Histones; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010472:Perchlorates; D010957:Plasmids; D013950:Thymus Gland; D014162:Transfection / Q000379:methods*
D002457:Cell Extracts; D003470:Culture Media; D006609:High Mobility Group Proteins; D006657:Histones; D010472:Perchlorates; D008156:Luciferases; D002118:Calcium