Submitted on September 13, 2005
Accepted on November 29, 2005
Deficiency of LKB1 in heart prevents ischemia-mediated activation of AMPK[alpha]2 but not AMPK[alpha]1
Kei Sakamoto1*, Elham Zarrinpashneh2, Grant R Budas3, Anne-Catherine Pouleur2, Anindya Dutta1, Alan R Prescott4, Jean-Louis Vanoverschelde2, Alan Ashworth5, Aleksandar Jovanovic3, Dario R Alessi1, and Luc Bertrand2
1 MRC Protein Phosphorylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom
2 Division of Cardiology, School of medicine, Universite catholique de Louvain, Brussels, Belgium
3 Maternal and Child Health Sciences, Tayside Institute of Child Health, Ninewells Hospital & Medical School, University of Dundee, Dundee, United Kingdom
4 Division of Cell Biology and Immunology, School of Life Sciences, University of Dundee, Dundee, United Kingdom
5 The Breakthrough Breast Cancer Research Centre, The Institute of Cancer Research, London, United Kingdom
* To whom correspondence should be addressed. E-mail: k.sakamoti{at}dundee.ac.uk.
Recent studies indicate that the LKB1 is a key regulator of the AMP-activated protein kinase (AMPK), which plays a crucial role in protecting cardiac muscle from damage during ischemia. We have employed mice that lack LKB1 in cardiac and skeletal muscle, and studied how this affected the activity of cardiac AMPK
1/2 under normoxic, ischemic and anoxic conditions. In the heart lacking cardiac muscle LKB1, the basal activity of AMPK2 was vastly reduced and not increased by ischemia or anoxia. Phosphorylation of AMPK2 at the site of LKB1 phosphorylation (Thr172) or phosphorylation of ACC2, a downstream substrate of AMPK, was ablated in ischemic heart lacking cardiac LKB1. Ischemia was found to increase the ADP/ATP and AMP/ATP ratios to a greater extent in LKB1-deficient cardiac muscle than in LKB1-expressing muscle. In contrast to AMPK2, significant basal activity of AMPK1 was observed in the lysates from the hearts lacking cardiac muscle LKB1, as well as in cardiomyocytes that had been isolated from these hearts. In the heart lacking cardiac LKB1, ischemia or anoxia induced a marked activation and phosphorylation of AMPK1, to a level that was only moderately lower than observed in LKB1-expressing heart. Echocardiographic and morphological analysis of the cardiac LKB1-deficient hearts indicated that these hearts were not overtly dysfunctional, despite possessing a reduced weight and enlarged atria. These findings indicate that LKB1 plays a crucial role in regulating AMPK2 activation, ACC2 phosphorylation and also regulating cellular energy levels in response to ischemia. They also provide genetic evidence that an alternative upstream kinase can activate AMPK1 in cardiac muscle.
