Host Pah1p phosphatidate phosphatase limits viral replication by regulating phospholipid synthesis.

Abstract

Replication of positive-strand RNA viruses [(+)RNA viruses] takes place in membrane-bound viral replication complexes (VRCs). Formation of VRCs requires virus-mediated manipulation of cellular lipid synthesis. Here, we report significantly enhanced brome mosaic virus (BMV) replication and much improved cell growth in yeast cells lacking PAH1 (pah1Δ), the sole yeast ortholog of human LIPIN genes. PAH1 encodes Pah1p (phosphatidic acid phosphohydrolase), which converts phosphatidate (PA) to diacylglycerol that is subsequently used for the synthesis of the storage lipid triacylglycerol. Inactivation of Pah1p leads to altered lipid composition, including high levels of PA, total phospholipids, ergosterol ester, and free fatty acids, as well as expansion of the nuclear membrane. In pah1Δ cells, BMV replication protein 1a and double-stranded RNA localized to the extended nuclear membrane, there was a significant increase in the number of VRCs formed, and BMV genomic replication increased by 2-fold compared to wild-type cells. In another yeast mutant that lacks both PAH1 and DGK1 (encodes diacylglycerol kinase converting diacylglycerol to PA), which has a normal nuclear membrane but maintains similar lipid compositional changes as in pah1Δ cells, BMV replicated as efficiently as in pah1Δ cells, suggesting that the altered lipid composition was responsible for the enhanced BMV replication. We further showed that increased levels of total phospholipids play an important role because the enhanced BMV replication required active synthesis of phosphatidylcholine, the major membrane phospholipid. Moreover, overexpression of a phosphatidylcholine synthesis gene (CHO2) promoted BMV replication. Conversely, overexpression of PAH1 or plant PAH1 orthologs inhibited BMV replication in yeast or Nicotiana benthamiana plants. Competing with its host for limited resources, BMV inhibited host growth, which was markedly alleviated in pah1Δ cells. Our work suggests that Pah1p promotes storage lipid synthesis and thus represses phospholipid synthesis, which in turn restricts both viral replication and cell growth during viral infection.

• Authors: Zhenlu Zhang, Guijuan He, Gil-Soo Han, Jiantao Zhang, Nicholas Catanzaro, Arturo Diaz, Zujian Wu, George M Carman, Lianhui Xie, Xiaofeng Wang
• Journal: PLoS pathogens (PLoS Pathog) Vol. 14 Issue 4 Pg. e1006988 (04 2018) ISSN: 1553-7374 [Electronic] United States
• PMID: 29649282 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Phospholipids
• Phosphatidate Phosphatase

Topics

• Bromovirus (physiology)
• Gene Expression Regulation, Fungal
• Gene Expression Regulation, Plant
• Genome, Viral
• Nuclear Envelope (metabolism)
• Phosphatidate Phosphatase (genetics, metabolism)
• Phospholipids (metabolism)
• Saccharomyces cerevisiae (metabolism, virology)
• Nicotiana (metabolism, virology)
• Virus Replication