Saccharomyces cerevisiae phospholipid:diacylglycerol acyl transferase (PDAT) devoid of its membrane anchor region is a soluble and active enzyme retaining its substrate specificities

Ghosal, Alokesh; Banaś, Antoni; Ståhl, Ulf; Dahlqvist, Anders; Lindqvist, Ylva; Stymne, Sten

doi:10.1016/j.bbalip.2007.10.007

Cited by 51 publications

(46 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The proposed critical role of PDAT1 in TAG synthesis is also consistent with the observation that overexpression of PDAT1, but not DGAT1, enhances TAG synthesis and LD formation in Arabidopsis leaves (Fan et al, 2013a(Fan et al, , 2013b, while disruption of PDAT1 decreases TAG accumulation with a simultaneous increase in membrane phospholipid levels in act1 and tgd1-1 (Fan et al, 2013a(Fan et al, , 2013b. In vitro assays showed that the PDAT enzyme catalyzes the transfer of acyl groups from a wide range of glycerolipid substrates to DAG to form TAG and lysolipids (Ståhl et al, 2004;Ghosal et al, 2007;Yoon et al, 2012), supporting the idea that PDAT1 may mediate the enhanced acyl flux through several major membrane lipid classes in the tgd1-1 mutant. However, it should be noted that PDAT1 and DGAT1 have overlapping functions in Arabidopsis ) and that DGAT1 is more important than PDAT1 in TAG synthesis at later stages of leaf development (Fan et al, 2013a(Fan et al, , 2013b, when the b-oxidation genes are coordinately induced (Troncoso-Ponce et al, 2013).…”

Section: Pdat1 and Sdp1 Function Cooperatively In Directing Fas Towarsupporting

confidence: 64%

Arabidopsis Lipins, PDAT1 Acyltransferase, and SDP1 Triacylglycerol Lipase Synergistically Direct Fatty Acids toward β-Oxidation, Thereby Maintaining Membrane Lipid Homeostasis

et al. 2014

View full text Add to dashboard Cite

Triacylglycerol (TAG) metabolism is a key aspect of intracellular lipid homeostasis in yeast and mammals, but its role in vegetative tissues of plants remains poorly defined. We previously reported that PHOSPHOLIPID:DIACYLGLYCEROL ACYLTRANSFERASE1 (PDAT1) is crucial for diverting fatty acids (FAs) from membrane lipid synthesis to TAG and thereby protecting against FA-induced cell death in leaves. Here, we show that overexpression of PDAT1 enhances the turnover of FAs in leaf lipids. Using the trigalactosyldiacylglycerol1-1 (tgd1-1) mutant, which displays substantially enhanced PDAT1-mediated TAG synthesis, we demonstrate that disruption of SUGAR-DEPENDENT1 (SDP1) TAG lipase or PEROXISOMAL TRANSPORTER1 (PXA1) severely decreases FA turnover, leading to increases in leaf TAG accumulation, to 9% of dry weight, and in total leaf lipid, by 3-fold. The membrane lipid composition of tgd1-1 sdp1-4 and tgd1-1 pxa1-2 double mutants is altered, and their growth and development are compromised. We also show that two Arabidopsis thaliana lipin homologs provide most of the diacylglycerol for TAG synthesis and that loss of their functions markedly reduces TAG content, but with only minor impact on eukaryotic galactolipid synthesis. Collectively, these results show that Arabidopsis lipins, along with PDAT1 and SDP1, function synergistically in directing FAs toward peroxisomal b-oxidation via TAG intermediates, thereby maintaining membrane lipid homeostasis in leaves.

show abstract

Section: Pdat1 and Sdp1 Function Cooperatively In Directing Fas Towarsupporting

confidence: 64%

Arabidopsis Lipins, PDAT1 Acyltransferase, and SDP1 Triacylglycerol Lipase Synergistically Direct Fatty Acids toward β-Oxidation, Thereby Maintaining Membrane Lipid Homeostasis

et al. 2014

View full text Add to dashboard Cite

show abstract

“…This is distinct from Sc-PDAT and At-PDAT, which exhibited higher substrate specificities toward PC and PE than the other PLs (Bana s et al, 2000; Dahlqvist et al, 2000;Ghosal et al, 2007). This difference can be explained in part by the distinct cellular PL compositions among yeasts, plants, and green algae.…”

Section: Cr-pdat Is a Multifunctional Enzymementioning

confidence: 94%

“…Previous studies showed an N-terminal deletion version of Sc-PDAT possessed low phospholipase activity in vitro (;2% of transacylation activity), which produced 2% free fatty acids during TAG production (Ghosal et al, 2007). At-PDAT has not been shown to have phospholipase activity Ståhl et al, 2004).…”

Section: Cr-pdat Lipase Activity With Broad Substrate Specificitymentioning

confidence: 99%

“…As shown in Figure 2A, both ORFCrPDAT and ΔTMCrPDAT exhibited a strong preference for anionic PL, including phosphatidic acid (PA), phosphatidylserine (PS), phosphatidylinositol (PI), and phosphatidylglycerol (PG), but showed less activities with the cationic PL (i.e., PC and phosphatidylethanolamine [PE]). Conversely, Sc-PDAT and At-PDAT exhibited strong preferences for PE and PC, respectively Ståhl et al, 2004;Ghosal et al, 2007). The enzymatic activity of DTMCrPDAT was determined using PI as an acyl donor and 1,2-DAG as an acyl acceptor.…”

Section: Typical Pdat Activity Of Cr-pdatmentioning

confidence: 99%

See 1 more Smart Citation

Phospholipid:Diacylglycerol Acyltransferase Is a Multifunctional Enzyme Involved in Membrane Lipid Turnover and Degradation While Synthesizing Triacylglycerol in the Unicellular Green Microalga Chlamydomonas reinhardtii

et al. 2012

View full text Add to dashboard Cite

Many unicellular microalgae produce large amounts (;20 to 50% of cell dry weight) of triacylglycerols (TAGs) under stress (e.g., nutrient starvation and high light), but the synthesis and physiological role of TAG are poorly understood. We present detailed genetic, biochemical, functional, and physiological analyses of phospholipid:diacylglycerol acyltransferase (PDAT) in the green microalga Chlamydomonas reinhardtii, which catalyzes TAG synthesis via two pathways: transacylation of diacylglycerol (DAG) with acyl groups from phospholipids and galactolipids and DAG:DAG transacylation. We demonstrate that PDAT also possesses acyl hydrolase activities using TAG, phospholipids, galactolipids, and cholesteryl esters as substrates. Artificial microRNA silencing of PDAT in C. reinhardtii alters the membrane lipid composition, reducing the maximum specific growth rate. The data suggest that PDAT-mediated membrane lipid turnover and TAG synthesis is essential for vigorous growth under favorable culture conditions and for membrane lipid degradation with concomitant production of TAG for survival under stress. The strong lipase activity of PDAT with broad substrate specificity suggests that this enzyme could be a potential biocatalyst for industrial lipid hydrolysis and conversion, particularly for biofuel production.

show abstract

“…However, PDAT was not considered as a significant source of acyl chains for Bayberry TAG because PC was not abundantly labeled, and PDAT expression was barely detectable (see RNA-seq results below). The labeling data were also not consistent with TAG synthesis by a DAG:DAG transacylase, as occurs in animal cells (Lehner and Kuksis, 1993;Yamashita et al, 2014), yeast (Ghosal et al, 2007), and safflower microsomes (Stobart et al, 1997). Specifically, a DAG:DAG transacylase would catalyze an exchange of acyl chains between two molecules of (Figure 2), which illustrated a lack of accumulation of glycerolipids within knob cells.…”

Section: Bayberry Surface Wax Is Produced By "Knobs" An Unusual Multmentioning

confidence: 76%

A Novel Pathway for Triacylglycerol Biosynthesis Is Responsible for the Accumulation of Massive Quantities of Glycerolipids in the Surface Wax of Bayberry (Myrica pensylvanica) Fruit

Simpson

Ohlrogge

2016

Plant Cell

View full text Add to dashboard Cite

ORCID ID: 0000-0003-4019-0681 (J.P.S.)Bayberry (Myrica pensylvanica) fruits synthesize an extremely thick and unusual layer of crystalline surface wax that accumulates to 32% of fruit dry weight, the highest reported surface lipid accumulation in plants. The composition is also striking, consisting of completely saturated triacylglycerol, diacylglycerol, and monoacylglycerol with palmitate and myristate acyl chains. To gain insight into the unique properties of Bayberry wax synthesis, we examined the chemical and morphological development of the wax layer, monitored wax biosynthesis through The most highly expressed lipid-related genes were associated with production of cutin, whereas transcripts for conventional TAG synthesis were >50-fold less abundant. The biochemical and expression data together indicate that Bayberry surface glycerolipids are synthesized by a pathway for TAG synthesis that is related to cutin biosynthesis. The combination of a unique surface wax and massive accumulation may aid understanding of how plants produce and secrete non-membrane glycerolipids and also how to engineer alternative pathways for lipid production in non-seeds.

show abstract

Saccharomyces cerevisiae phospholipid:diacylglycerol acyl transferase (PDAT) devoid of its membrane anchor region is a soluble and active enzyme retaining its substrate specificities

Cited by 51 publications

References 18 publications

Arabidopsis Lipins, PDAT1 Acyltransferase, and SDP1 Triacylglycerol Lipase Synergistically Direct Fatty Acids toward β-Oxidation, Thereby Maintaining Membrane Lipid Homeostasis

Arabidopsis Lipins, PDAT1 Acyltransferase, and SDP1 Triacylglycerol Lipase Synergistically Direct Fatty Acids toward β-Oxidation, Thereby Maintaining Membrane Lipid Homeostasis

Phospholipid:Diacylglycerol Acyltransferase Is a Multifunctional Enzyme Involved in Membrane Lipid Turnover and Degradation While Synthesizing Triacylglycerol in the Unicellular Green Microalga Chlamydomonas reinhardtii

A Novel Pathway for Triacylglycerol Biosynthesis Is Responsible for the Accumulation of Massive Quantities of Glycerolipids in the Surface Wax of Bayberry (Myrica pensylvanica) Fruit

Contact Info

Product

Resources

About