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Nucleic Acids Research, 1989, Vol. 17, No. 21 8783-8802
© 1989

MOLECULAR BIOLOGY

Identification of an amplification promoting DNA sequence from the hypotrichous cilate Stylonychia lemnae

Michael Wegner, Elke Helftenbein1, Friedemann Müller, Matthias Meinecke, Sabine Müller1 and Friedrich Grummt

Institut für Biochemie, Universität Würzburg D-8700 Wurzburg, FRG 1Biologie III, Abteilung Zellbiologie, Universität Tübingen D-7400 Tübingen, FRG

Received August 17, 1989. Revised October 3, 1989. Accepted October 3, 1989.

The macronucleus of the hypotrichous ciliate Stylonychia lemnae contains a 1218 bp long DNA molecule which becomes highly amplified during vegetative growth due to a continuous overreplication over a long time range. The region which is located upstream the open reading frame of the overamplified 1.2kbp Stylonychia DNA molecule enabled plasmids containing an inefficiently transcribed thymidine kinase gene to persist and amplify upon transfection into mouse L fibroblasts under selective conditions. This region contains long AT-rich stretches. The AT-rich sequences Interact with a previously characterized HMG-I like protein from mouse Ehrlich ascites tumour cells. A binding activity for AT-rich stretches could also be identified in macronuclear extracts from Stylonychia lemnae. We suggest a common mechanism for overamplification in Stylonychia macronuclei during vegetative growth and amplification of plasmid DNA in heterologous mouse cells under the influence of a common element.


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