Sophie Bamps1, Julia Wirtz, Fiona R Savory, Duncan Lake, Ian A Hope. 1. Institute of Integrative and Comparative Biology, Faculty of Biological Sciences, The University of Leeds, Woodhouse Lane, Leeds LS2 9JT, UK. s.bamps@leeds.ac.uk
Abstract
INTRODUCTION: As in yeast, flies and mammals, over-expression of the Caenorhabditis elegans sirtuin gene sir-2.1 leads to extension of lifespan and deletion of the gene shortens lifespan. The sir-2.1 gene, however, is located in an operon, an organization not taken into account in previous studies of this gene's expression. MATERIALS AND METHODS: Recombineering allowed insertion of both a mCherry and a gfp reporter gene precisely at the end of the two protein-coding regions of the 4.5kb sir-2.1 operon within a 29.3kb genomic DNA fosmid clone. RESULTS AND DISCUSSION: In C. elegans transgenic for this recombineered fosmid, with abundant food, the sir-2.1::mCherry distribution indicated that sir-2.1 is indeed expressed in the hypodermis and many nerve cells, as previously described, but also in the intestine and in muscles. This broader expression of sir-2.1, which would fit with an expectation that SIR2.1 function in influencing lifespan might be required in most cell types, arises from transcription starting with the gene upstream of sir-2.1 in the operon. Importantly, the expression of both genes in the operon increases upon starvation, this induction also depending on the operon promoter. Furthermore, SIR-2.1::mCherry undergoes a dynamic subcellular relocalization through starvation.
INTRODUCTION: As in yeast, flies and mammals, over-expression of the Caenorhabditis elegans sirtuin gene sir-2.1 leads to extension of lifespan and deletion of the gene shortens lifespan. The sir-2.1 gene, however, is located in an operon, an organization not taken into account in previous studies of this gene's expression. MATERIALS AND METHODS: Recombineering allowed insertion of both a mCherry and a gfp reporter gene precisely at the end of the two protein-coding regions of the 4.5kb sir-2.1 operon within a 29.3kb genomic DNA fosmid clone. RESULTS AND DISCUSSION: In C. elegans transgenic for this recombineered fosmid, with abundant food, the sir-2.1::mCherry distribution indicated that sir-2.1 is indeed expressed in the hypodermis and many nerve cells, as previously described, but also in the intestine and in muscles. This broader expression of sir-2.1, which would fit with an expectation that SIR2.1 function in influencing lifespan might be required in most cell types, arises from transcription starting with the gene upstream of sir-2.1 in the operon. Importantly, the expression of both genes in the operon increases upon starvation, this induction also depending on the operon promoter. Furthermore, SIR-2.1::mCherry undergoes a dynamic subcellular relocalization through starvation.
Authors: Ruben Nogueiras; Kirk M Habegger; Nilika Chaudhary; Brian Finan; Alexander S Banks; Marcelo O Dietrich; Tamas L Horvath; David A Sinclair; Paul T Pfluger; Matthias H Tschöp Journal: Physiol Rev Date: 2012-07 Impact factor: 37.312
Authors: Andreas H Ludewig; Yevgeniy Izrayelit; Donha Park; Rabia U Malik; Anna Zimmermann; Parag Mahanti; Bennett W Fox; Axel Bethke; Frank Doering; Donald L Riddle; Frank C Schroeder Journal: Proc Natl Acad Sci U S A Date: 2013-03-18 Impact factor: 11.205