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Characterization of major lipid droplet proteins from Dunaliella

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Abstract

Many green algal species can accumulate large amounts of triacylglycerides (TAG) under nutrient deprivation, making them a potential source for production of biodiesel. TAG are organized in cytoplasmic lipid bodies, which contain a major lipid droplet protein termed MLDP. Green algae MLDP differ in sequence from plant oleosins and from animal perilipins, and their structure and function are not clear. In this study, we describe the isolation of MLDP from three species of the extreme halotolerant green algae Dunaliella. Sequence alignment with other green algae MLDP proteins identified a conserved 4-proline domain that may be considered as a signature domain of Volvocales green algae MLDP. Gold immunolabeling localized MLDP at the surface of lipid droplets in D. salina. The induction of MLDP by nitrogen deprivation is kinetically correlated with TAG accumulation, and inhibition of TAG biosynthesis impairs MLDP accumulation suggesting that MLDP induction is co-regulated with TAG accumulation. These results can lead to a better understanding of the structure and function of Volvocales green algae MLDP proteins.

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Abbreviations

AA:

Amino acids

DMF:

Dimethylformamide

EDTA:

Ethylenediaminetetraacetic acid

GRAVY:

Grand average of hydropathy value

GSP:

Gene-specific primers

NGSP:

Nested gene-specific primers

KLH:

Keyhole limphet haemocyanine

MLDP:

Major lipid droplet protein

NR:

Nile red

PAGE:

Polyacrylamide gel electrophoresis

PAT:

Perilipin, adipophilin, TIP47, S3-12 and OXPAT

PBS:

Phosphate buffered saline

RACE:

Rapid amplification of the cDNA ends

RFU:

Relative fluorescence units

SDS:

Sodium dodecylsulfate

TAG:

Triacylglycerides

TLC:

Thin-layer chromatography

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Acknowledgments

We are grateful to the following scientists at The Weizmann Institute of Science: Mr. Vladimir Kiss from the Department of Biological Chemistry for his assistance in the Confocal Microscopy studies, Dr. Eyal Shimoni from the Department of Chemical Research Support for his help in the Electron Microscopy analyses and Dr. Miriam Eisenstein, also from the Department of Chemical Research Support, for her assistance and advice in the bioinformatic analysis. We are also grateful to Dr. Inna Khozin-Goldberg and to Prof. Zvi Cohen from The Ben-Gurion University for the GC–MS fatty acid analysis. This work was supported by The Alternative Energy Research Initiative Fund at The Weizmann Institute, by the Minerva Center: Photosynthesis under stress in changing world and by The Charles and Louise Gartner fund (to UP).

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  1. Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, 76100, Israel

    Lital Davidi, Adriana Katz & Uri Pick

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  1. Lital Davidi
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  2. Adriana Katz
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  3. Uri Pick
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Correspondence to Uri Pick.

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Davidi, L., Katz, A. & Pick, U. Characterization of major lipid droplet proteins from Dunaliella. Planta 236, 19–33 (2012). https://doi.org/10.1007/s00425-011-1585-7

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