Fatty Acid Transport in Saccharomyces cerevisiae

Zou, Zhiying; DiRusso, Concetta C.; Čtrnáctá, Vlasta; Black, Paul N.

doi:10.1074/jbc.m205034200

Cited by 110 publications

(67 citation statements)

References 50 publications

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“…For example, it is generally assumed that 10,16-dihydroxypalmitate and other cutin precursors are transferred through the plasma membrane (or an endomembrane) to the cell exterior. In many cases, cross-membrane transport of fatty acids requires an acyl-CoA synthetase activity on the distal side of the membrane (Schaffer and Lodish, 1994;Gargiulo et al, 1999;Zou et al, 2002), and acyl-CoAs have been shown to be substrates for cutin synthesis in vitro (Croteau and Kolattukudy, 1974). Thus, LACS2 may act either at the external face of the plasma membrane or the lumenal face of the ER or Golgi to synthesize hydroxyacyl-CoA substrates for cutin synthesis.…”

Section: Discussionmentioning

confidence: 99%

The Acyl-CoA Synthetase Encoded by LACS2 Is Essential for Normal Cuticle Development in Arabidopsis

2004

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Long-chain acyl-CoA synthetase (LACS) activities are encoded by a family of at least nine genes in Arabidopsis (Arabidopsis thaliana). These enzymes have roles in lipid synthesis, fatty acid catabolism, and the transport of fatty acids between subcellular compartments. Here, we show that the LACS2 gene (At1g49430) is expressed in young, rapidly expanding tissues, and in leaves expression is limited to cells of the adaxial and abaxial epidermal layers, suggesting that the LACS2 enzyme may act in the synthesis of cutin or cuticular waxes. A lacs2 null mutant was isolated by reverse genetics. Leaves of mutant plants supported pollen germination and released chlorophyll faster than wild-type leaves when immersed in 80% ethanol, indicating a defect in the cuticular barrier. The composition of surface waxes extracted from lacs2 leaves was similar to the wild type, and the total wax load was higher than the wild type (111.4 mg/dm 2 versus 76.4 mg/dm 2 , respectively). However, the thickness of the cutin layer on the abaxial surface of lacs2 leaves was only 22.3 6 1.7 nm compared with 33.0 6 2.0 nm for the wild type. In vitro assays showed that 16-hydroxypalmitate was an excellent substrate for recombinant LACS2 enzyme. We conclude that the LACS2 isozyme catalyzes the synthesis of v-hydroxy fatty acyl-CoA intermediates in the pathway to cutin synthesis. The lacs2 phenotype, like the phenotypes of some other cutin mutants, is very pleiotropic, causing reduced leaf size and plant growth, reduced seed production, and lower rates of seedling germination and establishment. The LACS2 gene and the corresponding lacs2 mutant will help in future studies of the cutin synthesis pathway and in understanding the consequences of reduced cutin production on many aspects of plant biology.

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Section: Discussionmentioning

confidence: 99%

The Acyl-CoA Synthetase Encoded by LACS2 Is Essential for Normal Cuticle Development in Arabidopsis

2004

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“…Using site-directed mutagenesis of the FATP1 orthologue in yeast Fat1p, fatty acid uptake and fatty acid activation are generally but not universally coupled processes (29). Four FAT1 alleles distinguish fatty acid transport and very long chain acyl-CoA synthetase activity, suggesting separable functions.…”

Section: Discussionmentioning

confidence: 99%

Characterization of the Acyl-CoA Synthetase Activity of Purified Murine Fatty Acid Transport Protein 1

Hall

Smith

Bernlohr

2003

Journal of Biological Chemistry

153

118

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Fatty acid transport protein 1 (FATP1) is an ϳ63-kDa plasma membrane protein that facilitates the influx of fatty acids into adipocytes as well as skeletal and cardiac myocytes. Previous studies with FATP1 expressed in COS1 cell extracts suggested that FATP1 exhibits very long chain acyl-CoA synthetase (ACS) activity and that such activity may be linked to fatty acid transport. To address the enzymatic activity of the isolated protein, murine FATP1 and ACS1 were engineered to contain a C-terminal Myc-His tag expressed in COS1 cells via adenoviral-mediated infection and purified to homogeneity using nickel affinity chromatography. Kinetic analysis of the purified enzymes was carried out for long chain palmitic acid (C16:0) and very long chain lignoceric acid (C24:0) as well as for ATP and CoA. FATP1 exhibited similar substrate specificity for fatty acids 16 -24 carbons in length, whereas ACS1 was 10-fold more active on long chain fatty acids relative to very long chain fatty acids. The very long chain acyl-CoA synthetase activity of the two enzymes was comparable as were the K m values for both ATP and coenzyme A. Interestingly, FATP1 was insensitive to inhibition by triacsin C, whereas ACS1 was inhibited by micromolar concentrations of the compound. These data represent the first characterization of purified FATP1 and indicate that the enzyme is a broad substrate specificity acyl-CoA synthetase. These findings are consistent with the hypothesis that that fatty acid uptake into cells is linked to their esterification with coenzyme A.

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“…Whereas it is highly speculative that the first 185 amino acids encoded by exons 1 and 2 would present a peptide with biological activity, it is worthwhile noting that this region is homologous to fragments of FATP1 that would include the entire extracellular N terminus, the transmembrane region, and cytoplasmic membraneassociated sequences (38). Because FATPs may be multifunctional proteins with separable residues involved in LCFA transport and activation (39), the hypothetical fusion proteins could retain some aspects of FATP4 function, such as LCFA transport, or interaction with other proteins, for instance longchain acyl-CoA synthetases (40) or other FATPs (19). Although the causes of neonatal mortality, described by Moulson et al (30) and Herrmann et al (22), may differ from the embryonic death described in this article, both lethalities ultimately underscore the importance of FATP4-mediated LCFA uptake/ activation for normal physiology.…”

Section: Table IIImentioning

confidence: 99%

Targeted Deletion of Fatty Acid Transport Protein-4 Results in Early Embryonic Lethality

Gimeno

Hirsch

Punreddy

et al. 2003

Journal of Biological Chemistry

127

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Fatty acid transport protein-4 (FATP4) is the major FATP in the small intestine. We previously demonstrated, using in vitro antisense experiments, that FATP4 is required for fatty acid uptake into intestinal epithelial cells. To further examine the physiological role of FATP4, mice carrying a targeted deletion of FATP4 were generated. Deletion of one allele of FATP4 resulted in 48% reduction of FATP4 protein levels and a 40% reduction of fatty acid uptake by isolated enterocytes. However, loss of one FATP4 allele did not cause any detectable effects on fat absorption on either a normal or a high fat diet. Deletion of both FATP4 alleles resulted in embryonic lethality as crosses between heterozygous FATP4 parents resulted in no homozygous offspring; furthermore, no homozygous embryos were detected as early as day 9.5 of gestation. Early embryonic lethality has been observed with deletion of other genes involved in lipid absorption in the small intestine, namely microsomal triglyceride transfer protein and apolipoprotein B, and has been attributed to a requirement for fat absorption early in embryonic development across the visceral endoderm. In mice, the extraembryonic endoderm supplies nutrients to the embryo prior to development of a chorioallantoic placenta. In wild-type mice we found that FATP4 protein is highly expressed by the epithelial cells of the visceral endoderm and localized to the brush-border membrane of extraembryonic endodermal cells. This localization is consistent with a role for FATP4 in fat absorption in early embryogenesis and suggests a novel requirement for FATP4 function during development.

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Fatty Acid Transport in Saccharomyces cerevisiae

Cited by 110 publications

References 50 publications

The Acyl-CoA Synthetase Encoded by LACS2 Is Essential for Normal Cuticle Development in Arabidopsis

The Acyl-CoA Synthetase Encoded by LACS2 Is Essential for Normal Cuticle Development in Arabidopsis

Characterization of the Acyl-CoA Synthetase Activity of Purified Murine Fatty Acid Transport Protein 1

Targeted Deletion of Fatty Acid Transport Protein-4 Results in Early Embryonic Lethality

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