Two transgenic mouse models for β-subunit components of succinate-CoA ligase yielding pleiotropic metabolic alterations.

Gergely Kacsó ,Kimi Araki,Christos Chinopoulos,Ory Madgar,Chaya Miller,Zsuzsanna Vargedo,Beáta Németh,Masatake Araki,Haruka Ito,Ann Saada,Polina Ilin,Mai Nakahara,Zoltán Nagy ,Attila Patócs ,Vera Ádám-Vizi ,I Iordanov ,Anikó Gál ,Mária Judit Molnár ,Judit Dóczi ,Daniel J Adams ,Elsebet Østergaard ,Dóra Ravasz

doi:10.1042/bcj20160594

Abstract

Succinate-CoA ligase (SUCL) is a heterodimer enzyme composed of Suclg1 α-subunit and a substrate-specific Sucla2 or Suclg2 β-subunit yielding ATP or GTP, respectively. In humans, the deficiency of this enzyme leads to encephalomyopathy with or without methylmalonyl aciduria, in addition to resulting in mitochondrial DNA depletion. We generated mice lacking either one Sucla2 or Suclg2 allele. Sucla2 heterozygote mice exhibited tissue- and age-dependent decreases in Sucla2 expression associated with decreases in ATP-forming activity, but rebound increases in cardiac Suclg2 expression and GTP-forming activity. Bioenergetic parameters including substrate-level phosphorylation (SLP) were not different between wild-type and Sucla2 heterozygote mice unless a submaximal pharmacological inhibition of SUCL was concomitantly present. mtDNA contents were moderately decreased, but blood carnitine esters were significantly elevated. Suclg2 heterozygote mice exhibited decreases in Suclg2 expression but no rebound increases in Sucla2 expression or changes in bioenergetic parameters. Surprisingly, deletion of one Suclg2 allele in Sucla2 heterozygote mice still led to a rebound but protracted increase in Suclg2 expression, yielding double heterozygote mice with no alterations in GTP-forming activity or SLP, but more pronounced changes in mtDNA content and blood carnitine esters, and an increase in succinate dehydrogenase activity. We conclude that a partial reduction in Sucla2 elicits rebound increases in Suclg2 expression, which is sufficiently dominant to overcome even a concomitant deletion of one Suclg2 allele, pleiotropically affecting metabolic pathways associated with SUCL. These results as well as the availability of the transgenic mouse colonies will be of value in understanding SUCL deficiency.

Highlights

Succinate-CoA ligase (SUCL), known as succinyl coenzyme A synthetase, or succinate thiokinase is a heterodimer enzyme composed of an invariant α-subunit encoded by SUCLG1 and a substratespecific β-subunit encoded by either SUCLA2 or SUCLG2
In Sucla2 heterozygote mice, there is a rebound increase in Suclg2 expression associated with mostly unaffected bioenergetic parameters, while mtDNA contents are moderately decreased in some organs, and blood carnitine levels are elevated
Insertion of the retroviral vector into the Sucla2 gene led to the splicing of the endogenous upstream exons into this cassette to produce a fusion that leads to termination of further transcription of the endogenous Sucla2 exons downstream of the insertion

Summary

Introduction

Succinate-CoA ligase (SUCL), known as succinyl coenzyme A synthetase, or succinate thiokinase is a heterodimer enzyme composed of an invariant α-subunit encoded by SUCLG1 and a substratespecific β-subunit encoded by either SUCLA2 or SUCLG2. In the tissues of Sucla heterozygote mice, we investigated respiration rates and membrane potential (ΔΨm) for an array of mitochondrial substrates and various metabolic states, complex I, II, II/III and IV activities, as well as SLP during respiratory inhibition or true anoxia. We cross-bred Sucla2+/− with Suclg2+/− mice, yielding double heterozygote Sucla2+/−/Suclg2+/− mice, and investigated the expression of g1, g2 and a2 subunits, mtDNA content, blood carnitine esters and bioenergetic parameters. In Sucla heterozygote mice, there is a rebound increase in Suclg expression associated with mostly unaffected bioenergetic parameters, while mtDNA contents are moderately decreased in some organs, and blood carnitine levels are elevated. Results obtained from embryonic tissues of Sucla2−/− mice have been published in ref. [18] and are discussed in relation to the results obtained here

Experimental procedures

Results

Discussion