Identification of Dicarboxylate Carrier Slc25a10 as Malate Transporter in de Novo Fatty Acid Synthesis

Mitochondrial solute carrier family 25 member 10 (Slc25a10) transports dicarboxylates such as malate or succinate across the mitochondrial inner membrane. Although fatty acid synthesis in adipose tissue or the liver is initiated by citrate transport in exchange for malate across the mitochondrial membrane, the transporter responsible for supplying malate during citrate transport has not been identified. In the present study, we clarified the role of Slc25a10 in supplying malate for citrate transport and examined the effect of Slc25a10 suppression on the lipogenic pathway and lipid accumulation. We have reported an Slc25a10 increase in white adipose tissue in obese mouse models and a decrease in a fasted mouse model using expression profiles. Next, we examined the effect of Slc25a10 suppression by small interfering RNA on citrate transport in the lipogenic cell lines HepG2 and 3T3-L1. We observed that inhibition of malate transport by Slc25a10 suppression significantly reduced the citrate transport from the mitochondria to the cytosol. We also found that suppression of Slc25a10 down-regulated the lipogenic pathway, indicated by decreases in ACC1 expression and suppression of Slc25a10 lipid in that Slc25a10 an role in supplying malate for citrate transport for fatty acid synthesis and indicated that inhibition of Slc25a10 lipid in adipose Mitochondrial solute carrier family 25 member 10 (Slc25a10) transports dicarboxylates such as malate or succinate across the mitochondrial inner membrane. Although fatty acid synthesis in adipose tissue or the liver is initiated by citrate transport in exchange for malate across the mitochondrial membrane, the transporter responsible for supplying malate during citrate transport has not been identified. In the present study, we clarified the role of Slc25a10 in supplying malate for citrate transport and examined the effect of Slc25a10 suppression on the lipogenic pathway and lipid accumulation. We have reported an Slc25a10 increase in white adipose tissue in obese mouse models and a decrease in a fasted mouse model using expression profiles. Next, we examined the effect of Slc25a10 suppression by small interfering RNA on citrate transport in the lipogenic cell lines HepG2 and 3T3-L1. We observed that inhibition of malate transport by Slc25a10 suppression significantly reduced the citrate transport from the mitochondria to the cytosol. We also found that suppression of Slc25a10 down-regulated the lipogenic pathway, indicated by decreases in ACC1 expression and suppression of Slc25a10 lipid in that Slc25a10 an role in supplying malate for citrate transport for fatty acid synthesis and indicated that inhibition of Slc25a10 lipid in adipose fatty acid synthesis in lipogenic such as white adipose tissue white adipose fatty acid solute carrier family 25 member small interfering white adipose fatty acid solute carrier family 25 member small interfering and the of fatty acid synthesis the of mitochondrial citrate to the in exchange for malate by a citrate carrier by the of citrate to by citrate is to by or the of fatty acid synthesis citrate also as an for the of fatty acid fatty by by fatty acid and fatty acid or of fatty acid such as or in the of In the of fatty acid synthesis from the is the to the transport of citrate to the in the of fatty acid the the of malate for the citrate the malate and mitochondrial to solute carrier family 25 member 10 transports a such as across the mitochondrial inner membrane. carrier is of and is to the inner mitochondrial Slc25a10 is in lipogenic expression in and and expression in the liver expression is also in the of is in lipogenic and for citrate of Slc25a10 in in of mitochondria an the of Slc25a10 in fatty acid synthesis has not been have been to or in the of the from the of We also the that is in of obese a and of lipogenic obese and models or in the of and in the pathway of fatty acid and synthesis have as suppression of the expression or of and obese in models of RNA suppression in that the expression of reduced in and of in pathway the present study, we a of using obese mouse models and a fasted mouse model to on in the lipogenic Slc25a10 in obese mouse models and down-regulated in the mouse We examined the effect of Slc25a10 suppression on fatty acid We found that Slc25a10 a role in supplying malate for the citrate transport for fatty acid We also found that the suppression of Slc25a10 down-regulated fatty acid in reduced lipid in and in of and and of and obese and have been In the fasted fasted for and to a for of and by the and in RNA from using and an in 10 of RNA for of of to for the and of using the RNA from and as in the the for of and to for of and using an expression to or and for in for and for of HepG2 the a of for Slc25a10 using as by in an a we of a in of for HepG2 the the the of and for mouse Slc25a10 by as a of in of for the the for the of mitochondria from HepG2 malate by in 10 of a transport and in the of and and to the carrier and citrate mitochondria the transport for mitochondria and in of the transport of exchange using the mitochondria in of and of transport initiated by and the indicated by the of mitochondria by and and using a of and of malate in malate from and using a of and in cell in by the of acid cell by the in malate malate and to and of by of using the for in citrate 10 citrate citrate malate and to malate and of by of malate and citrate in the mitochondria of in from in the of and using a of a from in the to a and to and using a by the of to of the and for in of in an of and to the for and of for 10 to and by in of cell by for of on and for in and the of the and by by of and as using a of in and the an lipogenic pathway in we in of obese and fasted mouse obese mouse models and and and the fasted mouse model a is to on the of is to increase in by the of by we by in the obese models and down-regulated by in the fasted on the in is adipose the is in the obese mouse model and in fasted the mitochondrial transporter of Slc25a10 is in and is also an for citrate across the mitochondrial membrane. We on that the carrier is in the of fatty acid in and fasted of in the obese model and of in fasted carrier family 25 member in a Next, we the expression of Slc25a10 in obese mouse models by and observed in and We also a increase in Slc25a10 in of Slc25a10 in the of Slc25a10 in and the effect of inhibition on fatty acid we for and mouse In lipogenic from liver and mouse for Slc25a10 we observed and of suppression in HepG2 and Next, we the of Slc25a10 in mitochondria from the suppression of Slc25a10 expression by mitochondria from HepG2 and and the malate exchange reduced to and in of HepG2 and of of Slc25a10 malate transport in suppression of expression by in HepG2 and expression by malate transport across the mitochondrial inner in mitochondria from or HepG2 or expression of citrate and carrier by of for Slc25a10 by the expression of solute carrier citrate and the of to Slc25a10 not by citrate carrier transport in mitochondria also as a exchange of the citrate carrier not by Slc25a10 suppression of Slc25a10 and We examined the effect of Slc25a10 suppression on malate and citrate in of Slc25a10 in HepG2 malate in the malate by Slc25a10 transports malate across the inner of mitochondrial malate also to the in cell malate from the mitochondria or cytosol. malate in malate and mitochondria observed that malate in the of by the inhibition of malate transport from the mitochondria to the cytosol. citrate carrier transports malate in exchange for mitochondrial we that the reduced malate by Slc25a10 suppression decrease exchange of citrate citrate In the citrate of by the in mitochondrial of Slc25a10 malate and citrate in cytosol. HepG2 or in cell and mitochondrial in or in cell and in mitochondrial of Slc25a10 ACC1 from is from as an of the effect of reduced citrate on we the expression of ACC1 in the suppression of Slc25a10 in by Although Slc25a10 in of Slc25a10 suppression in for Slc25a10 10 of ACC1 also in on that the lipogenic pathway in In ACC1 reduced to of in for Slc25a10 10 of We also the expression of and citrate carrier as and in and in and expression of lipogenic also citrate and in not by and In lipogenic cell reduced ACC1 expression also observed for Slc25a10 citrate carrier expression not expression of ACC1 for Slc25a10 in and in In Slc25a10 in Slc25a10 ACC1 expression significantly in citrate carrier expression not of Slc25a10 decreases ACC1 and or the expression of Slc25a10 ACC1 citrate carrier citrate and to by In HepG2 or ACC1 and citrate carrier also by expression of in by in HepG2 and the expression of In the of in citrate is to citrate by Slc25a10 suppression increase the of expression of in and of ACC1 and expression in HepG2 or the expression of and ACC1 during of or in ACC1 in we in and from and to fatty acid in of fatty acid the of In HepG2 and of and in observed and is in fatty acid synthesis by or indicated that reduced decreases the of fatty acid of Slc25a10 decreases and an of fatty in HepG2 and of Slc25a10 in to fatty acid such as citrate ACC1 and the effect of Slc25a10 suppression on the of fatty acid the of lipid in in the or the of the lipid of the of lipid synthesis reduced by in the effect of Slc25a10 suppression on lipid we the of in for Slc25a10 by lipid observed in In and the of the lipid We lipid by and the In a increase in lipid observed in In lipid reduced by in of Slc25a10 decreases lipid in in by of lipid in and during lipid in and and by clarified the role of Slc25a10 in the of fatty acid Although the of Slc25a10 in has been is the that Slc25a10 is for citrate transport across the mitochondrial inner membrane. we that the inhibition of Slc25a10 down-regulated to reduced lipid of Slc25a10 ACC1 expression and the of ACC1 by Slc25a10 to We that the reduced citrate by Slc25a10 suppression ACC1 citrate is an for to of ACC1 of the a of ACC1 fatty acid synthesis in lipid accumulation. reduced or ACC1 expression a ACC1 is by fatty by fatty acid synthesis ACC1 expression in that citrate in the reduced in the role of the carrier in citrate transport for fatty acid is to reduced exchange of citrate and malate across the mitochondrial membrane. citrate carrier citrate the for fatty acid synthesis from the mitochondrial in exchange for malate We that Slc25a10 malate the citrate carrier for fatty acid synthesis by mitochondrial malate the cytosol. is that reduced malate by Slc25a10 suppression decrease the exchange of citrate carrier in the of fatty acid in reduced lipid accumulation. is also by tissue or that that malate have on an effect of malate transport on the to reduced to transport from the mitochondrial to the cytosol. In the malate is from citrate by the of citrate and malate malate is to by the and is mitochondria for to In obese the malate by Slc25a10 the the of citrate to the of the and and and or to the from the by also in and suppression of a transporter by RNA to a increase in and decreases in and is to suppression of the reduced of the of In reduced is to the decrease in citrate for fatty acid synthesis by the transporter that suppression of or the of citrate for fatty acid an to lipid by Slc25a10 suppression that Slc25a10 a for an In inhibition of Slc25a10 in is to the carrier is in adipose tissue in to lipogenic in a of Slc25a10 has been an the to the and we that Slc25a10 an role in supplying malate for the citrate transport for fatty acid synthesis and that inhibition of Slc25a10 fatty acid to lipid accumulation. of the of Slc25a10 in to effect of suppression is fatty acid synthesis in lipogenic such as white adipose tissue white adipose fatty acid solute carrier family 25 member small interfering white adipose fatty acid solute carrier family 25 member small interfering and the of fatty acid synthesis the of mitochondrial citrate to the in exchange for malate by a citrate carrier by the of citrate to by citrate is to by or the of fatty acid synthesis citrate also as an for the of fatty acid fatty by by fatty acid and fatty acid or of fatty acid such as or in the of In the of fatty acid synthesis from the is the to the transport of citrate to the in the of fatty acid the the of malate for the citrate the malate and mitochondrial to Mitochondrial solute carrier family 25 member 10 transports a such as across the mitochondrial inner membrane. carrier is of and is to the inner mitochondrial Slc25a10 is in lipogenic expression in and and expression in the liver expression is also in the of is in lipogenic and for citrate of Slc25a10 in in of mitochondria an the of Slc25a10 in fatty acid synthesis has not been have been to or in the of the from the of We also the that is in of obese a and of lipogenic obese and models or in the of and in the pathway of fatty acid and synthesis have as suppression of the expression or of and obese in models of RNA suppression in that the expression of reduced in and of in pathway In the present study, we a of using obese mouse models and a fasted mouse model to on in the lipogenic Slc25a10 in obese mouse models and down-regulated in the mouse We examined the effect of Slc25a10 suppression on fatty acid We found that Slc25a10 a role in supplying malate for the citrate transport for fatty acid We also found that the suppression of Slc25a10 down-regulated fatty acid in reduced lipid in and in of and and of and obese and have been In the fasted fasted for and to a for of and by the and in RNA from using and an in 10 of RNA for of of to for the and of using the RNA from and as in the the for of and to for of and using an expression to or and for in for and for of HepG2 the a of for Slc25a10 using as by in an a we of a in of for HepG2 the the the of and for mouse Slc25a10 by as a of in of for the the for the of mitochondria from HepG2 malate by in 10 of a transport and in the of and and to the carrier and citrate mitochondria the transport for mitochondria and in of the transport of exchange using the mitochondria in of and of transport initiated by and the indicated by the of mitochondria by and and using a of and of malate in malate from and using a of and in cell in by the of acid cell by the in malate malate and to and of by of using the for in citrate 10 citrate citrate malate and to malate and of by of malate and citrate in the mitochondria of in from in the of and using a of a from in the to a and to and using a by the of to of the and for in of in an of and to the for and of for 10 to and by in of cell by for of on and for in and the of the and by by of and as using a and in of and and of and obese and have been In the fasted fasted for and to a for of and by the and in RNA from using and an in 10 of RNA for of of to for the and of using the RNA from and as in the the for of and to for of and using an expression to or and for in for and for of HepG2 the a of for Slc25a10 using as by in an a we of a in of for HepG2 the the the of and for mouse Slc25a10 by as a of in of for the the for the of Mitochondrial mitochondria from HepG2 malate by in 10 of a transport and in the of and and to the carrier and citrate mitochondria the transport for mitochondria and in of the transport of exchange using the mitochondria in of and of transport initiated by and the indicated by the of mitochondria by and and using a of and of malate in malate from and using a of and in cell in by the of acid cell by the in malate malate and to and of by of using the for in citrate 10 citrate citrate malate and to malate and of by of malate and citrate in the mitochondria of in from in the of and using a of a from in the to a and to and using a by the of to of the and for in of in an of and to the for and of for 10 to and by in of cell by for of on and for in and the of the and by by of and as using a of in and the an lipogenic pathway in we in of obese and fasted mouse obese mouse models and and and the fasted mouse model a is to on the of is to increase in by the of by we by in the obese models and down-regulated by in the fasted on the in is adipose the is in the obese mouse model and in fasted the mitochondrial transporter of Slc25a10 is in and is also an for citrate across the mitochondrial membrane. We on that the carrier is in the of fatty acid in and fasted of in the obese model and of in fasted carrier family 25 member in a Next, we the expression of Slc25a10 in obese mouse models by and observed in and We also a increase in Slc25a10 in of Slc25a10 in the of Slc25a10 in and the effect of inhibition on fatty acid we for and mouse In lipogenic from liver and mouse for Slc25a10 we observed and of suppression in HepG2 and Next, we the of Slc25a10 in mitochondria from the suppression of Slc25a10 expression by mitochondria from HepG2 and and the malate exchange reduced to and in of HepG2 and of of for Slc25a10 by the expression of solute carrier citrate and the of to Slc25a10 not by citrate carrier transport in mitochondria also as a exchange of the citrate carrier not by Slc25a10 suppression of Slc25a10 and We examined the effect of Slc25a10 suppression on malate and citrate in of Slc25a10 in HepG2 malate in the malate by Slc25a10 transports malate across the inner of mitochondrial malate also to the in cell malate from the mitochondria or cytosol. malate in malate and mitochondria observed that malate in the of by the inhibition of malate transport from the mitochondria to the cytosol. citrate carrier transports malate in exchange for mitochondrial we that the reduced malate by Slc25a10 suppression decrease exchange of citrate citrate In the citrate of by the in mitochondrial of Slc25a10 malate and citrate in cytosol. HepG2 or in cell and mitochondrial in or in cell and in mitochondrial of Slc25a10 ACC1 from is from as an of the effect of reduced citrate on we the expression of ACC1 in the suppression of Slc25a10 in by Although Slc25a10 in of Slc25a10 suppression in for Slc25a10 10 of ACC1 also in on that the lipogenic pathway in In ACC1 reduced to of in for Slc25a10 10 of We also the expression of and citrate carrier as and in and in and expression of lipogenic also citrate and in not by and In lipogenic cell reduced ACC1 expression also observed for Slc25a10 citrate carrier expression not expression of ACC1 for Slc25a10 in and in In Slc25a10 in Slc25a10 ACC1 expression significantly in citrate carrier expression not of Slc25a10 decreases ACC1 and or the expression of Slc25a10 ACC1 citrate carrier citrate and to by In HepG2 or ACC1 and citrate carrier also by expression of in by in HepG2 and the expression of In the of in citrate is to citrate by Slc25a10 suppression increase the of expression of in and of ACC1 and expression in HepG2 or the expression of and ACC1 during of or in ACC1 in we in and from and to fatty acid in of fatty acid the of In HepG2 and of and in observed and is in fatty acid synthesis by or indicated that reduced decreases the of fatty acid of Slc25a10 decreases and an of fatty in HepG2 and of Slc25a10 in to fatty acid such as citrate ACC1 and the effect of Slc25a10 suppression on the of fatty acid the of lipid in in the or the of the lipid of the of lipid synthesis reduced by in the effect of Slc25a10 suppression on lipid we the of in for Slc25a10 by lipid observed in In and the of the lipid We lipid by and the In a increase in lipid observed in In lipid reduced by in of Slc25a10 decreases lipid in in by of lipid in and during lipid in and and by of in and the an lipogenic pathway in we in of obese and fasted mouse obese mouse models and and and the fasted mouse model a is to on the of is to increase in by the of by we by in the obese models and down-regulated by in the fasted on the in is adipose the is in the obese mouse model and in fasted the mitochondrial transporter of Slc25a10 is in and is also an for citrate across the mitochondrial membrane. We on that the carrier is in the of fatty acid Next, we the expression of Slc25a10 in obese mouse models by and observed in and We also a increase in Slc25a10 in of Slc25a10 in the of Slc25a10 in and the effect of inhibition on fatty acid we for and mouse In lipogenic from liver and mouse for Slc25a10 we observed and of suppression in HepG2 and Next, we the of Slc25a10 in mitochondria from the suppression of Slc25a10 expression by mitochondria from HepG2 and and the malate exchange reduced to and in of HepG2 and of of for Slc25a10 by the expression of solute carrier citrate and the of to Slc25a10 not by citrate carrier transport in mitochondria also as a exchange of the citrate carrier not by Slc25a10 suppression of Slc25a10 and We examined the effect of Slc25a10 suppression on malate and citrate in of Slc25a10 in HepG2 malate in the malate by Slc25a10 transports malate across the inner of mitochondrial malate also to the in cell malate from the mitochondria or cytosol. malate in malate and mitochondria observed that malate in the of by the inhibition of malate transport from the mitochondria to the cytosol. citrate carrier transports malate in exchange for mitochondrial we that the reduced malate by Slc25a10 suppression decrease exchange of citrate citrate In the citrate of by the in mitochondrial of Slc25a10 ACC1 from is from as an of the effect of reduced citrate on we the expression of ACC1 in the suppression of Slc25a10 in by Although Slc25a10 in of Slc25a10 suppression in for Slc25a10 10 of ACC1 also in on that the lipogenic pathway in In ACC1 reduced to of in for Slc25a10 10 of We also the expression of and citrate carrier as and in and in and expression of lipogenic also citrate and in not by and In lipogenic cell reduced ACC1 expression also observed for Slc25a10 citrate carrier expression not expression of ACC1 for Slc25a10 in and in In Slc25a10 in Slc25a10 ACC1 expression significantly in citrate carrier expression not expression of in by in HepG2 and the expression of In the of in citrate is to citrate by Slc25a10 suppression increase the of in ACC1 in we in and from and to fatty acid in of fatty acid the of In HepG2 and of and in observed and is in fatty acid synthesis by or indicated that reduced decreases the of fatty acid of Slc25a10 in to fatty acid such as citrate ACC1 and the effect of Slc25a10 suppression on the of fatty acid the of lipid in in the or the of the lipid of the of lipid synthesis reduced by in the effect of Slc25a10 suppression on lipid we the of in for Slc25a10 by lipid observed in In and the of the lipid We lipid by and the In a increase in lipid observed in In lipid reduced by in clarified the role of Slc25a10 in the of fatty acid Although the of Slc25a10 in has been is the that Slc25a10 is for citrate transport across the mitochondrial inner membrane. we that the inhibition of Slc25a10 down-regulated to reduced lipid of Slc25a10 ACC1 expression and the of ACC1 by Slc25a10 to We that the reduced citrate by Slc25a10 suppression ACC1 citrate is an for to of ACC1 of the a of ACC1 fatty acid synthesis in lipid accumulation. reduced or ACC1 expression a ACC1 is by fatty by fatty acid synthesis ACC1 expression in that citrate in the reduced in the role of the carrier in citrate transport for fatty acid is to reduced exchange of citrate and malate across the mitochondrial membrane. citrate carrier citrate the for fatty acid synthesis from the mitochondrial in exchange for malate We that Slc25a10 malate the citrate carrier for fatty acid synthesis by mitochondrial malate the cytosol. is that reduced malate by Slc25a10 suppression decrease the exchange of citrate carrier in the of fatty acid in reduced lipid accumulation. is also by tissue or that that malate have on an effect of malate transport on the to reduced to transport from the mitochondrial to the cytosol. In the malate is from citrate by the of citrate and malate malate is to by the and is mitochondria for to In obese the malate by Slc25a10 the the of citrate to the of the and and and or to the from the by also in and suppression of a transporter by RNA to a increase in and decreases in and is to suppression of the reduced of the of In reduced is to the decrease in citrate for fatty acid synthesis by the transporter that suppression of or the of citrate for fatty acid an to lipid by Slc25a10 suppression that Slc25a10 a for an In inhibition of Slc25a10 in is to the carrier is in adipose tissue in to lipogenic in a of Slc25a10 has been an the to the and we that Slc25a10 an role in supplying malate for the citrate transport for fatty acid synthesis and that inhibition of Slc25a10 fatty acid to lipid accumulation. of the of Slc25a10 in to effect of suppression is We clarified the role of Slc25a10 in the of fatty acid Although the of Slc25a10 in has been is the that Slc25a10 is for citrate transport across the mitochondrial inner membrane. we that the inhibition of Slc25a10 down-regulated to reduced lipid accumulation. of Slc25a10 ACC1 expression and the of ACC1 by Slc25a10 to We that the reduced citrate by Slc25a10 suppression ACC1 citrate is an for to of ACC1 of the a of ACC1 fatty acid synthesis in lipid accumulation. reduced or ACC1 expression a ACC1 is by fatty by fatty acid synthesis ACC1 expression in We that citrate in the reduced in the role of the carrier in citrate transport for fatty acid is to reduced exchange of citrate and malate across the mitochondrial membrane. citrate carrier citrate the for fatty acid synthesis from the mitochondrial in exchange for malate We that Slc25a10 malate the citrate carrier for fatty acid synthesis by mitochondrial malate the cytosol. is that reduced malate by Slc25a10 suppression decrease the exchange of citrate carrier in the of fatty acid in reduced lipid accumulation. is also by tissue or that that malate have on an effect of malate transport on the to reduced to transport from the mitochondrial to the cytosol. In the malate is from citrate by the of citrate and malate malate is to by the and is mitochondria for to In obese the malate by Slc25a10 the the of citrate to the of the In and and and or to the from the by also in and suppression of a transporter by RNA to a increase in and decreases in and is to suppression of the reduced of the of In reduced is to the decrease in citrate for fatty acid synthesis by the transporter that suppression of or the of citrate for fatty acid an to lipid by Slc25a10 suppression that Slc25a10 a for an In inhibition of Slc25a10 in is to the carrier is in adipose tissue in to lipogenic in a of Slc25a10 has been an the to the and In we that Slc25a10 an role in supplying malate for the citrate transport for fatty acid synthesis and that inhibition of Slc25a10 fatty acid to lipid accumulation. of the of Slc25a10 in to effect of suppression is