DNA strand-transfer activity in pea (Pisum sativum L.) chloroplasts

Heriberto D Cerutti, André T. Jagendorf

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The occurrence of DNA recombination in plastids of higher plants is well documented. However, little is known at the enzymic level. To begin dissecting the biochemical mechanism(s) involved we focused on a key step: strand transfer between homologous parental DNAs. We detected a RecA-like strand transfer activity in stromal extracts from pea (Pisum sativum L.) chloroplasts. Formation of joint molecules requires Mg2+, ATP, and homologous substrates. This activity is inhibited by excess single-stranded DNA (ssDNA), suggesting a necessary stoichiometric relation between enzyme and ssDNA. In a novel assay with Triton X-100-permeabilized chloroplasts, we also detected strand invasion of the endogenous chloroplast DNA by 32P-labeled ssDNA complementary to the 16S rRNA gene. Joint molecules, analyzed by electron microscopy, contained the expected displacement loops.

Original languageEnglish (US)
Pages (from-to)145-153
Number of pages9
JournalPlant Physiology
Volume102
Issue number1
DOIs
StatePublished - Jan 1 1993

Fingerprint

single-stranded DNA
Single-Stranded DNA
Peas
Chloroplasts
Pisum sativum
peas
chloroplasts
DNA
Joints
Chloroplast DNA
Plastids
biochemical mechanisms
Octoxynol
chloroplast DNA
rRNA Genes
Genetic Recombination
plastids
electron microscopy
Electron Microscopy
Adenosine Triphosphate

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science

Cite this

DNA strand-transfer activity in pea (Pisum sativum L.) chloroplasts. / Cerutti, Heriberto D; Jagendorf, André T.

In: Plant Physiology, Vol. 102, No. 1, 01.01.1993, p. 145-153.

Research output: Contribution to journalArticle

Cerutti, Heriberto D ; Jagendorf, André T. / DNA strand-transfer activity in pea (Pisum sativum L.) chloroplasts. In: Plant Physiology. 1993 ; Vol. 102, No. 1. pp. 145-153.
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