Production of RNA by a polymerase protein encapsulated within phospholipid vesicles.

Abstract:

:Catalyzed polymerization reactions represent a primary anabolic activity of all cells. It can be assumed that early cells carried out such reactions, in which macromolecular catalysts were encapsulated within some type of boundary membrane. In the experiments described here, we show that a template-independent RNA polymerase (polynucleotide phosphorylase) can be encapsulated in dimyristoyl phosphatidylcholine vesicles without substrate. When the substrate adenosine diphosphate (ADP) was provided externally, long-chain RNA polymers were synthesized within the vesicles. Substrate flux was maximized by maintaining the vesicles at the phase transition temperature of the component lipid. A protease was introduced externally as an additional control. Free enzyme was inactivated under identical conditions. RNA products were visualized in situ by ethidium bromide fluorescence. The products were harvested from the liposomes, radiolabeled, and analyzed by polyacrylamide gel electrophoresis. Encapsulated catalysts represent a model for primitive cellular systems in which an RNA polymerase was entrapped within a protected microenvironment.

journal_name

J Mol Evol

authors

Chakrabarti AC,Breaker RR,Joyce GF,Deamer DW

doi

10.1007/BF00160400

subject

Has Abstract

pub_date

1994-12-01 00:00:00

pages

555-9

issue

6

eissn

0022-2844

issn

1432-1432

journal_volume

39

pub_type

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