Planta

, Volume 219, Issue 2, pp 325–331 | Cite as

Functional analysis and regulation of the malate synthase from Chlamydomonas reinhardtii

Original Article

Abstract

Malate synthase (EC 2.3.3.9, formerly EC 4.1.2.2) has been investigated in the unicellular green algae Chlamydomonas reinhardtii. The molecular characteristics and the regulation of gene expression have been investigated for the enzyme. A full-length malate synthase cDNA has been isolated, containing an open reading frame of 1,641 bp encoding a polypeptide of 546 amino acids. This protein shares the conserved signature of the malate synthase family, along with the catalytic residues essential for enzymatic activity and a C-terminal motif that matches the consensus for glyoxysome import. Functionality studies have been facilitated by heterologous expression of the malate synthase cDNA in Escherichia coli. The remarkable metabolic versatility of the alga has been used to analyse the metabolic control of malate synthase gene expression. The data strongly support the role of acetate and light as the main regulatory effectors, and the existence of cross-talk between the two signalling pathways.

Keywords

Chlamydomonas Glyoxylate cycle Malate synthase 

Abbreviations

IPTG

Isopropyl β-d-thiogalactopyranoside

MS

Malate synthase

PCR

Polymerase chain reaction

PTS

Peroxisomal targeting sequence

RACE

Rapid amplification of cDNA ends

TAP

Tris–acetate–phosphate medium

TCA

Tricarboxylic acid cycle

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • J. Nogales
    • 2
  • M. I. Guijo
    • 1
  • A. Quesada
    • 1
  • F. Merchán
    • 1
  1. 1.Department of Biochemistry and Molecular Biology, Faculty of Veterinary MedicineUniversity of ExtremaduraCáceresSpain
  2. 2.Department of Molecular MicrobiologyCentro de Investigaciones Biológicas-CSICMadridSpain

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