DOI: 10.5061/dryad.gs7s3
title: Data from: Are Niemann-Pick type C proteins key players in cnidarian-dinoflagellate endosymbioses?

description: The symbiotic interaction between cnidarians, such as corals and sea
 anemones, and the unicellular algae Symbiodinium is regulated by yet
 poorly understood cellular mechanisms, despite the ecological importance
 of coral reefs. These mechanisms, including host-symbiont recognition and
 metabolic exchange, control symbiosis stability under normal conditions,
 but also lead to symbiosis breakdown (bleaching) during stress. This study
 describes the repertoire of the sterol-trafficking proteins Niemann-Pick
 type C (NPC1 and NPC2) in the symbiotic sea anemone Anemonia viridis. We
 found one NPC1 gene instead of two in vertebrates. While only one NPC2
 gene is present in most metazoans, this gene has been duplicated in
 cnidarians and we detected four NPC2 genes in A. viridis. However, only
 one gene (AvNPC2-d) was upregulated in symbiotic sea anemones and
 displayed higher expression in the gastrodermis (symbiont-containing
 tissue) than in the epidermis. We performed immunolabeling experiments on
 tentacle cross sections and demonstrated that the AvNPC2-d protein was
 closely associated with symbiosomes. In addition, AvNPC1 and AvNPC2-d gene
 expression was strongly downregulated during stress, especially at the
 onset of symbiosis breakdown. These data suggest that AvNPC2-d is involved
 in both the stability and dysfunction of cnidarian-dinoflagellate
 symbioses.
description: Alignment file of NPC2
 protein sequences_Figure1ASequences of NPC2
 proteins aligned using the MAFFT program. This alignment was used to
 construct a phylogenetic tree using the maximum likelihood method
 (phyML3.0 software) and the WAG+I+G substitution model (I=0.057 and gamma
 factor=5.831). Accession number of sequences: AvNPC2-d,HG670301; AvNPC2-c,
 HG670300; NvNPC2-2, XP_001627355; NvNPC2-3, XP_001635502; AdNPC2-2,
 adi_v1.06781; AdNPC2-3, adi_v1.23434; AdNPC2-4, adi_v1.10342; AvNPC2-b,
 HG670299; AdNPC2-1, adi_v1.14987 + contig adi_v1.08570; AdNPC2-5,
 aug_v2a.08569; AdNPC2-6, aug_v2a.08570; AvNPC2-a, HG670298; NvNPC2-1,
 XP_001622874; HsNPC2, NP_006423.1; DrNPC2-a, NP_001122191.1; DrNPC2-b,
 NP_775331.1; CiNPC2-1, XP_002127695.1; TaNPC2, XP_002109765.1; HvNPC2-3,
 XP_002166396.2; Hv NPC2-4, XP_002166374.2; Hv NPC2-2, XP_002164043.1; Hv
 NPC2-1, XP_002156875.1; AqNPC2, CL5422Contig1 from AQUE_CAP3_100104
 assembly from compagenAlignment file of NPC1
 protein sequences_Figure2Sequences of NPC1
 proteins aligned using the MAFFT program. This alignment was used to
 generate a phylogenetic tree using the phyML 3.0 sofware, the maximum
 likelihood method and the LG+I+G substitution model (I=0.1 and gamma
 factor=1.266). Accession numbers: HsNPC1L1, NP_001095118.1; BtNPC1L1,
 XP_588051; DrNPC1L1, XP_002663230.2; CiNPC1-b, XP_002122922; DmNPC1-a,
 NP_609357; DmNPC1-b, NP_608417; HsNPC1, AAK25791.1; BtNPC1, AAI51277;
 DrNPC1, XP_001919958.3; CiNPC1-a, XP_002120129; SpNPC1, XP_780036; AvNPC1,
 HG670297; NvNPC1, XM_001634881.1; AdNPC1, adi_v1.19271 from
 ADIG_G-PEP_111201 database; HmNPC1, XM_002164383.1; AqNPC1,
 XP_003384854.1;PrimersEfficiencyPCR efficiency of primers used in this study (DNA quantification and heat stress experiment)PrimerEfficiency.xlsxqPCR_QuantifZxAvSymbiodinium quantification by real-time quantitative PCR measurement of the relative nuclear gene copy number (symbiont to host nuclei) during hyperthermal stress.qPCR_HeatStressRelative gene expression data analysis (raw Ct and normalized expression data) for the heat stress experimentNormalizationfactor_HeatStressNormalization factor was calculated based on the expression level of RCC2 and COP-ɣ genes using the GeNorm software, for the heat stress experiment.TreeFile_NPC2_Figure1Tree data generated using the maximum likelihood method and the PhyML3.0 software. Compute these data with the SeaView program to display the NPC2 phylogenetic tree.TreeFile_NPC1_Figure2Tree data generated using the maximum likelihood method and the PhyML3.0 software. Compute these data with the SeaView program to display the NPC1 phylogenetic tree.