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Perilipin-related protein regulates lipid metabolism in C. elegans.

Chughtai AA, Kaššák F, Kostrouchová M, Novotný JP, Krause MW, Saudek V, Kostrouch Z, Kostrouchová M - PeerJ (2015)

Bottom Line: Perilipins have been identified in organisms as diverse as metazoa, fungi, and amoebas but strikingly not in nematodes.In contrast to embryos, lipid-containing structures in enterocytes and in epidermal cells of adult animals are smaller in mutants than in wild type animals.Our results demonstrate the existence of a perilipin-related regulation of fat metabolism in nematodes and provide new possibilities for functional studies of lipid metabolism.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Cellular Biology and Pathology, First Faculty of Medicine, Charles University in Prague , Albertov, Prague , Czech Republic.

ABSTRACT
Perilipins are lipid droplet surface proteins that contribute to fat metabolism by controlling the access of lipids to lipolytic enzymes. Perilipins have been identified in organisms as diverse as metazoa, fungi, and amoebas but strikingly not in nematodes. Here we identify the protein encoded by the W01A8.1 gene in Caenorhabditis elegans as the closest homologue and likely orthologue of metazoan perilipin. We demonstrate that nematode W01A8.1 is a cytoplasmic protein residing on lipid droplets similarly as human perilipins 1 and 2. Downregulation or elimination of W01A8.1 affects the appearance of lipid droplets resulting in the formation of large lipid droplets localized around the dividing nucleus during the early zygotic divisions. Visualization of lipid containing structures by CARS microscopy in vivo showed that lipid-containing structures become gradually enlarged during oogenesis and relocate during the first zygotic division around the dividing nucleus. In mutant embryos, the lipid containing structures show defective intracellular distribution in subsequent embryonic divisions and become gradually smaller during further development. In contrast to embryos, lipid-containing structures in enterocytes and in epidermal cells of adult animals are smaller in mutants than in wild type animals. Our results demonstrate the existence of a perilipin-related regulation of fat metabolism in nematodes and provide new possibilities for functional studies of lipid metabolism.

No MeSH data available.


Related in: MedlinePlus

Identification of C. elegans protein W01A8.1a as a close homologue of vertebrate perilipin.C. elegans protein W01A8.1a is compared with nematode homologues of pairwise sequence identity lower then 70% and with Plin2 and 3 from two diverse vertebrates. The three perilipin specific domains (indicated in red) were identified through homology with human Plin3. The six 11-mer repeats in W01A8.1a (positions 126–136, 137–147, 148–158, 159–169, 170–180 and 181–191) were established with HHrepID algorithm (Biegert & Soding, 2008). The N-terminal PAT domain is thought to interact with HSL. The central domain consisting of imperfect 11-mer repeats forming amphipathic helices is responsible for the main affinity to LDs and the C-terminal domain containing an apolipoprotein-like 4-helix bundle probably plays an additional role in the affinity to LDs and is known to interact with ABHD5 in mammalian Plin1 and 3 (Brasaemle, 2007). Alignment was done using T-coffee alignment of all available nematode sequences aligned with vertebrate Plin2 and 3 sequences in three iterations using ProfileAlign routine in MyHits suite (myhits.isb-sib.ch). Selected sequences from top to bottom: (Species, database identifier): Caenorhabditis elegans, Q23095; Strongyloides ratti, CACX01001972.1; Loa loa, E1G5Y0 and ADBU02007219.1; Haemonchus contortus, CDJ80228.1; Bursaphelenchus xylophilus, CADV01008520.1; Heterorhabditis bacteriophora, ES742365.1 and ACKM01001830.1; Ascaris suum, U1NU60; Homo sapiens 2, PLIN2_HUMAN; Homo sapiens 3, PLIN3_HUMAN; Latimeria chalumnae 2, H3AYC0; Latimeria chalumnae 3, GAAA01019375.1. Nucleotide sequences were translated with Wise2 program (Birney, Clamp & Durbin, 2004). Amino acid types are colored according to the Clustal scheme (jalview.org/help/html/colourSchemes/clustal.html).
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fig-1: Identification of C. elegans protein W01A8.1a as a close homologue of vertebrate perilipin.C. elegans protein W01A8.1a is compared with nematode homologues of pairwise sequence identity lower then 70% and with Plin2 and 3 from two diverse vertebrates. The three perilipin specific domains (indicated in red) were identified through homology with human Plin3. The six 11-mer repeats in W01A8.1a (positions 126–136, 137–147, 148–158, 159–169, 170–180 and 181–191) were established with HHrepID algorithm (Biegert & Soding, 2008). The N-terminal PAT domain is thought to interact with HSL. The central domain consisting of imperfect 11-mer repeats forming amphipathic helices is responsible for the main affinity to LDs and the C-terminal domain containing an apolipoprotein-like 4-helix bundle probably plays an additional role in the affinity to LDs and is known to interact with ABHD5 in mammalian Plin1 and 3 (Brasaemle, 2007). Alignment was done using T-coffee alignment of all available nematode sequences aligned with vertebrate Plin2 and 3 sequences in three iterations using ProfileAlign routine in MyHits suite (myhits.isb-sib.ch). Selected sequences from top to bottom: (Species, database identifier): Caenorhabditis elegans, Q23095; Strongyloides ratti, CACX01001972.1; Loa loa, E1G5Y0 and ADBU02007219.1; Haemonchus contortus, CDJ80228.1; Bursaphelenchus xylophilus, CADV01008520.1; Heterorhabditis bacteriophora, ES742365.1 and ACKM01001830.1; Ascaris suum, U1NU60; Homo sapiens 2, PLIN2_HUMAN; Homo sapiens 3, PLIN3_HUMAN; Latimeria chalumnae 2, H3AYC0; Latimeria chalumnae 3, GAAA01019375.1. Nucleotide sequences were translated with Wise2 program (Birney, Clamp & Durbin, 2004). Amino acid types are colored according to the Clustal scheme (jalview.org/help/html/colourSchemes/clustal.html).

Mentions: We performed BLASTp searches with individual protein sequences of human perilipins that generated no significant hits to Nematoda sequences in the UniProt database, consistent with previous efforts that failed to identify a perilipin-related protein in this phylum. However, when a sequence alignment of chordate perilipins 2 and 3 (OMA database) was submitted as query in PSI-BLAST, the C. elegans protein W01A8.1a (Q23095_CAEEL) was identified as a highly significant hit (E = 3 × 10−13). A reciprocal PSI-BLAST search with the aligned closest nematode homologues of W01A8.1a identified chordate perilipins as strong hits with human Perilipin 2 (significance score E = 10−53) appearing in the second iteration of the search. Similarly, HHpred profile-to-profile searches with human perilipin sequences as a query of the C. elegans proteome identified proteins coded by W01A8.1 (a, b or c) and reciprocally W01A8.1a showed profile homology to all human perilipins and the corresponding Pfam (Punta et al., 2012) perilipin profile (PF03036). Each available nematode proteome contained only a single such perilipin-related sequence, in stark contrast to the insect and chordate proteomes that had 2–5 perilipin paralogues. A sequence alignment of Plin2 and 3 from two selected vertebrates is compared with their nematode homologues (Fig. 1). Although the sequence-to sequence comparisons are not sufficient to unravel the sequence homology between vertebrate and nematode Plins, the similarity appear clearly in the profile-to sequence (PSI-BLAST) and profile-to-profile (HHpred) searches. We conclude that vertebrate Plins and nematode W01A8.1 are statistically highly significant homologues.


Perilipin-related protein regulates lipid metabolism in C. elegans.

Chughtai AA, Kaššák F, Kostrouchová M, Novotný JP, Krause MW, Saudek V, Kostrouch Z, Kostrouchová M - PeerJ (2015)

Identification of C. elegans protein W01A8.1a as a close homologue of vertebrate perilipin.C. elegans protein W01A8.1a is compared with nematode homologues of pairwise sequence identity lower then 70% and with Plin2 and 3 from two diverse vertebrates. The three perilipin specific domains (indicated in red) were identified through homology with human Plin3. The six 11-mer repeats in W01A8.1a (positions 126–136, 137–147, 148–158, 159–169, 170–180 and 181–191) were established with HHrepID algorithm (Biegert & Soding, 2008). The N-terminal PAT domain is thought to interact with HSL. The central domain consisting of imperfect 11-mer repeats forming amphipathic helices is responsible for the main affinity to LDs and the C-terminal domain containing an apolipoprotein-like 4-helix bundle probably plays an additional role in the affinity to LDs and is known to interact with ABHD5 in mammalian Plin1 and 3 (Brasaemle, 2007). Alignment was done using T-coffee alignment of all available nematode sequences aligned with vertebrate Plin2 and 3 sequences in three iterations using ProfileAlign routine in MyHits suite (myhits.isb-sib.ch). Selected sequences from top to bottom: (Species, database identifier): Caenorhabditis elegans, Q23095; Strongyloides ratti, CACX01001972.1; Loa loa, E1G5Y0 and ADBU02007219.1; Haemonchus contortus, CDJ80228.1; Bursaphelenchus xylophilus, CADV01008520.1; Heterorhabditis bacteriophora, ES742365.1 and ACKM01001830.1; Ascaris suum, U1NU60; Homo sapiens 2, PLIN2_HUMAN; Homo sapiens 3, PLIN3_HUMAN; Latimeria chalumnae 2, H3AYC0; Latimeria chalumnae 3, GAAA01019375.1. Nucleotide sequences were translated with Wise2 program (Birney, Clamp & Durbin, 2004). Amino acid types are colored according to the Clustal scheme (jalview.org/help/html/colourSchemes/clustal.html).
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Related In: Results  -  Collection

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Show All Figures
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fig-1: Identification of C. elegans protein W01A8.1a as a close homologue of vertebrate perilipin.C. elegans protein W01A8.1a is compared with nematode homologues of pairwise sequence identity lower then 70% and with Plin2 and 3 from two diverse vertebrates. The three perilipin specific domains (indicated in red) were identified through homology with human Plin3. The six 11-mer repeats in W01A8.1a (positions 126–136, 137–147, 148–158, 159–169, 170–180 and 181–191) were established with HHrepID algorithm (Biegert & Soding, 2008). The N-terminal PAT domain is thought to interact with HSL. The central domain consisting of imperfect 11-mer repeats forming amphipathic helices is responsible for the main affinity to LDs and the C-terminal domain containing an apolipoprotein-like 4-helix bundle probably plays an additional role in the affinity to LDs and is known to interact with ABHD5 in mammalian Plin1 and 3 (Brasaemle, 2007). Alignment was done using T-coffee alignment of all available nematode sequences aligned with vertebrate Plin2 and 3 sequences in three iterations using ProfileAlign routine in MyHits suite (myhits.isb-sib.ch). Selected sequences from top to bottom: (Species, database identifier): Caenorhabditis elegans, Q23095; Strongyloides ratti, CACX01001972.1; Loa loa, E1G5Y0 and ADBU02007219.1; Haemonchus contortus, CDJ80228.1; Bursaphelenchus xylophilus, CADV01008520.1; Heterorhabditis bacteriophora, ES742365.1 and ACKM01001830.1; Ascaris suum, U1NU60; Homo sapiens 2, PLIN2_HUMAN; Homo sapiens 3, PLIN3_HUMAN; Latimeria chalumnae 2, H3AYC0; Latimeria chalumnae 3, GAAA01019375.1. Nucleotide sequences were translated with Wise2 program (Birney, Clamp & Durbin, 2004). Amino acid types are colored according to the Clustal scheme (jalview.org/help/html/colourSchemes/clustal.html).
Mentions: We performed BLASTp searches with individual protein sequences of human perilipins that generated no significant hits to Nematoda sequences in the UniProt database, consistent with previous efforts that failed to identify a perilipin-related protein in this phylum. However, when a sequence alignment of chordate perilipins 2 and 3 (OMA database) was submitted as query in PSI-BLAST, the C. elegans protein W01A8.1a (Q23095_CAEEL) was identified as a highly significant hit (E = 3 × 10−13). A reciprocal PSI-BLAST search with the aligned closest nematode homologues of W01A8.1a identified chordate perilipins as strong hits with human Perilipin 2 (significance score E = 10−53) appearing in the second iteration of the search. Similarly, HHpred profile-to-profile searches with human perilipin sequences as a query of the C. elegans proteome identified proteins coded by W01A8.1 (a, b or c) and reciprocally W01A8.1a showed profile homology to all human perilipins and the corresponding Pfam (Punta et al., 2012) perilipin profile (PF03036). Each available nematode proteome contained only a single such perilipin-related sequence, in stark contrast to the insect and chordate proteomes that had 2–5 perilipin paralogues. A sequence alignment of Plin2 and 3 from two selected vertebrates is compared with their nematode homologues (Fig. 1). Although the sequence-to sequence comparisons are not sufficient to unravel the sequence homology between vertebrate and nematode Plins, the similarity appear clearly in the profile-to sequence (PSI-BLAST) and profile-to-profile (HHpred) searches. We conclude that vertebrate Plins and nematode W01A8.1 are statistically highly significant homologues.

Bottom Line: Perilipins have been identified in organisms as diverse as metazoa, fungi, and amoebas but strikingly not in nematodes.In contrast to embryos, lipid-containing structures in enterocytes and in epidermal cells of adult animals are smaller in mutants than in wild type animals.Our results demonstrate the existence of a perilipin-related regulation of fat metabolism in nematodes and provide new possibilities for functional studies of lipid metabolism.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Cellular Biology and Pathology, First Faculty of Medicine, Charles University in Prague , Albertov, Prague , Czech Republic.

ABSTRACT
Perilipins are lipid droplet surface proteins that contribute to fat metabolism by controlling the access of lipids to lipolytic enzymes. Perilipins have been identified in organisms as diverse as metazoa, fungi, and amoebas but strikingly not in nematodes. Here we identify the protein encoded by the W01A8.1 gene in Caenorhabditis elegans as the closest homologue and likely orthologue of metazoan perilipin. We demonstrate that nematode W01A8.1 is a cytoplasmic protein residing on lipid droplets similarly as human perilipins 1 and 2. Downregulation or elimination of W01A8.1 affects the appearance of lipid droplets resulting in the formation of large lipid droplets localized around the dividing nucleus during the early zygotic divisions. Visualization of lipid containing structures by CARS microscopy in vivo showed that lipid-containing structures become gradually enlarged during oogenesis and relocate during the first zygotic division around the dividing nucleus. In mutant embryos, the lipid containing structures show defective intracellular distribution in subsequent embryonic divisions and become gradually smaller during further development. In contrast to embryos, lipid-containing structures in enterocytes and in epidermal cells of adult animals are smaller in mutants than in wild type animals. Our results demonstrate the existence of a perilipin-related regulation of fat metabolism in nematodes and provide new possibilities for functional studies of lipid metabolism.

No MeSH data available.


Related in: MedlinePlus