Characterisation of the gene family encoding acetoacetyl-CoA thiolase in Arabidopsis
Iván Ahumada A C , Albert Cairó A , Andréa Hemmerlin B , Víctor González A D , Irene Pateraki A , Thomas J. Bach B , Manuel Rodríguez-Concepción A D , Narciso Campos A and Albert Boronat A EA Departament de Bioquímica i Biologia Molecular, Facultat de Biologia, Universitat de Barcelona, Avda. Diagonal 645, 08028 Barcelona, Spain.
B Centre National de la Recherche Scientifique, UPR 2357, Institut de Biologie Moléculaire des Plantes, 28 rue Goethe, 67083 Strasbourg Cedex, France.
C Present address: Universidad Autónoma de Chile-Sede Talca, Escuela de Ciencias de la Salud, 5 Poniente Número 1670, Talca, Chile.
D Present address: Centre de Recerca en Agrigenòmica (CRAG-CSIC), Jordi Girona 18-26, 08034 Barcelona, Spain.
E Corresponding author. Email: aboronat@ub.edu
Functional Plant Biology 35(11) 1100-1111 https://doi.org/10.1071/FP08012
Submitted: 19 January 2008 Accepted: 30 July 2008 Published: 28 November 2008
Abstract
Thiolases are ubiquitous enzymes involved in many essential biochemical processes. Biosynthetic thiolases, also known as acetoacetyl-CoA thiolases (AACT), catalyse a reversible Claisen-type condensation of two acetyl-CoA molecules to form acetoacetyl-CoA. Here, we report the characterisation of two genes from Arabidopsis thaliana L., ACT1 and ACT2, which encode two closely related AACT isoforms (AACT1 and AACT2, respectively). Transient expression of constructs encoding AACT1 and AACT2 fused to GFP revealed that the two proteins show a different subcellular localisation. While AACT1 is found in peroxisomes, AACT2 localises in the cytosol and the nucleus. The peroxisomal localisation of AACT1 depends on the presence of a C-terminal peroxisomal targeting sequence (PTS1) motif (Ser-Ala-Leu) not previously found in other organisms. ACT1 and ACT2 genes are also differentially expressed. Whereas ACT2 is expressed at relatively high level in all plant tissues, the expression of ACT1 is restricted to roots and inflorescences and its transcript is present at very low levels. The obtained results are in agreement with the involvement of AACT2 in catalysing the first step of the mevalonate pathway. The metabolic function of AACT1 is not clear at present, although its particular peroxisomal localisation might exclude a role in isoprenoid biosynthesis.
Additional keywords: acetyl-CoA, isoprenoid biosynthesis, mevalonate pathway, peroxisome.
Acknowledgements
This work was supported by grants from the Spanish Ministerio de Educación y Ciencia (BIO2006–03704 to A.B., BFU2006–14655 to N.C. and BIO2005–00367 to M.R.C., all including FEDER funds), and the Generalitat de Catalunya (grant 2005SGR-00914). This work has been carried out within the Centre CONSOLIDER on Agrigenomics funded by the Spanish Ministry of Education and Science. I.A. was recipient of a pre-doctoral MUTIS fellowship from the Agencia Española de Cooperación International. We thank Aurora Piñas and Antonio Comaposada for their contribution to the isolation and preliminary characterisation of ACT2 cDNA. We also thank Dr Julio Rozas for his advice in the phylogenetic analysis of thiolases and the staff from the Serveis de Camps Experimentals and Serveis Cientificotecnics of the Universitat de Barcelona for their help in plant culture, DNA-sequencing and confocal laser scanning microscopy.
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