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Partial purification and characterization of the mRNA complementing a temperature-sensitive S-phase cell cycle mutation.

A Fainsod·M Marcus·P F Lin·F H Ruddle
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Abstract

E36ts24 is a temperature-sensitive (ts) cell cycle mutant derived from the Chinese hamster lung cell line E36. At the restrictive temperature for growth (40.3 degrees C), the mutant cells are arrested at early S phase. We have microinjected poly(A)+ RNA isolated from the wild-type E36 cells into the cytoplasm of E36ts24 cells arrested at early S phase after 24 hr of incubation at 40.3 degrees C. The ts mutation was transiently complemented in a significant fraction of the microinjected cells as evidenced by the incorporation of [3H]thymidine assayed by autoradiography. Microinjection of mRNA fractionated by methylmercuric hydroxide/agarose gel showed that the mRNA capable of transiently complementing the mutation in E36ts24 contains about 940 nucleotides. Hence, it can code at most for a protein containing about 230 amino acids. We estimate that the partial purification by fractionation of the mRNA active in the transient complementation is on the order of 100- to 200-fold.

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AnimalsCell LineCricetinaeCricetulusGenetic Complementation TestInterphaseLungMolecular WeightMutationPoly ARNARNA, MessengerTemperatureThymidine KinaseJournal ArticleResearch Support, U.S. Gov't, Non-P.H.S.

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A Fainsod, M Marcus, P F Lin, F H Ruddle. 1984. Partial purification and characterization of the mRNA complementing a temperature-sensitive S-phase cell cycle mutation.. https://doi.org/10.1073/pnas.81.8.2393

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