Molecular Mechanism of Citrate Efflux by the Mitochondrial Citrate Transporter CT in Filamentous Fungus WJ11

Overview

Journal Front Microbiol

Specialty Microbiology

Date 2021 May 31

PMID 34054781

Citations 7

Authors

Wu Yang

Shiqi Dong

Junhuan Yang

Hassan Mohamed

Aabid Manzoor Shah

Yusuf Nazir

Xiuzhen Gao

Huirong Fan

Yuanda Song

Affiliations

Soon will be listed here.

Abstract

The mitochondrial citrate transporter (MCT) plays an important role in citrate efflux from the mitochondria in eukaryotes, and hence provides a direct correlation between carbohydrate metabolism and lipid synthesis. Our previous studies on transporters confirmed the presence of two MCTs (TCT and CT) in oleaginous WJ11 associated with high lipid accumulation. However, the molecular mechanism of citrate efflux from the mitochondria by MCT in is still unclear. To study the citrate transport mechanism of CT, the citrate transporter gene was expressed in , and its product was purified. The citrate transport activity of the protein was studied in CT reconstituted liposomes. Our results showed high efficiency of CT for [C] citrate/citrate exchange with 0.01 mM at 25°C. Besides citrate, other molecules such as oxaloacetate, malate, fumarate, succinate aconitate, oxoadipate, isocitrate, and glutamate also promote citrate transport. In addition, the overexpression and knockout plasmids were constructed and transferred into WJ11, and the mitochondria were isolated, and the transport activity was studied. Our findings showed that in the presence of 10 mM malate, the mitochondria of -overexpressing transformant showed 51% increase in the efflux rate of [C] citrate, whereas the mitochondria of the -knockout transformant showed 18% decrease in citrate efflux compared to the mitochondria of wild-type WJ11. This study provided the first mechanistic evidence of citrate efflux from the mitochondria by citrate transporter in oleaginous filamentous fungus , which is associated with high lipid accumulation.

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