Table 1.
Neuropeptide/peptide hormones predicted on the basis of transcriptome mining of Bugula neritina.
Fig 1.
Precursor sequences of neuropeptide/peptide hormones from Bugula neritina.
Green: signal peptide; Red: putative mature peptide; Blue: preprotein sequence; Black underline: dibasic cleavage site.
Table 2.
Predicted neuropeptide/peptide hormones from Bugula neritina.
Fig 2.
A) Primary structure of the deduced precursor sequence of BnFDSIG-like and BnPedal/orcokinin-like and B) multiple sequence alignment of the mature FDSIG and Pedal/orcokinin neuropeptides.
Mature peptide sequences were derived from precursor protein sequences download from NCBI GenBank. PdFDSIG-P1-3: (AAP57098.1) pedal peptide preprohormone [Lymnaea stagnalis]; CrNLP-14-P1-3: (EFO91279.1) CRE-NLP-14 protein [Caenorhabditis remanei]; AsOrcokininB-P1-3: (ERG84279.1) orcokinin peptides type b [Ascaris suum]; CgOrcokinin-P1-3: (XP_011447518.1) PREDICTED: orcokinin peptides type A-like [Crassostrea gigas]; AcPedal-1-P1-3: (NP_001191585.1) pedal peptide-1 precursor [Aplysia californica]; LsPedal-P1-2: (AAP57098.1) pedal peptide preprohormone [Lymnaea stagnalis]; AaOrcokinin-P1-3: (AFK81939.1) orcokinin [Amphibalanus amphitrite]; BmOrcokininP1-2: BAG50371.1| orcokinin [Bombyx mori].
Fig 3.
Expression of BnFDSIG-like, BnILP-B, and BnGHB5 during the larval stage.
(A) Whole-mount in situ hybridization (WISH), (B) a representative section of WISH and C) enlarged image of the region highlighted by the red dotted line in B), which includes the apical organ and neural plexus.
Fig 4.
Sequence alignment of the Bugula neritina putative mature glycoprotein hormone beta 5 with homologs.
CgGHB5: (EKC24490.1) glycoprotein hormone beta 5 Crassostrea gigas; TcGHB5: (EFA10760.1glycoprotein hormone beta 5) Tribolium castaneum; TnGHB5: (KRZ59516.1) glycoprotein hormone beta 5 Trichinella nativa; BmGHB5: (NP_001124380.1) glycoprotein hormone beta 5 Bombyx mori; DrGHB5: (XP_003198997.1) glycoprotein hormone beta 5 Danio rerio; LaGHB5-1 (XP_013400795.1) glycoprotein hormone beta 5-like isoform 1 Lingula anatina; LaGHB5-2: (XP_013400794.1) glycoprotein hormone beta 5-like isoform 2 Lingula anatina; DmGHB5: (NP_001104335.1) glycoprotein hormone beta 5 Drosophila melanogaster; HsGHB5: (AAO33390.1) glycoprotein hormone beta 5. Homo sapiens. The asterisk “*” marks the conserved cysteine that will form dimers.
Fig 5.
Sequence alignment of Bugula neritina putative mature Neuropeptide F with homologs.
AcNPY: (NP_001191635.1) pro-neuropeptide Y precursor Aplysia californica; HsNPYY: (CAG46926.1) NPYY Homo sapiens; LsNPY: (CAB63265.1) NPY Lymnaea stagnalis; NvNPFL: (XP_008206766) NPF Nasonia vitripennis; DmBPYY: (AGB96023.1) NPF Drosophila melanogaster; BmNPY: (NP_001124361.1) NPF Bombyx mori. The RXR(F/Y) modified are conserved except in the putative mature peptide of BnNPY. The asterisk “*” denotes the conservation of tyrosine (Y) residues that are predicted to be sulfated in the mature peptide in the NPF family.
Fig 6.
Sequence alignment of the Bugula neritina insulin-like peptide precursor protein with homologs.
CgMIRP-3-like: (XP_011415724.1) PREDICTED: molluscan insulin-related peptide 3-like [Crassostrea gigas]; BgMIRP-3-like: (XP_013070273.1) PREDICTED: molluscan insulin-related peptide 3-like [Biomphalaria glabrata]; CgILP: (XP_011455161.1) PREDICTED: insulin-like peptide [Crassostrea gigas]; AcIns: (AF160192_1) insulin precursor [Aplysia californica]; PdILP 3: (AHB62360.1) insulin-related peptide 3 neuropeptide precursor [Platynereis dumerilii]; PdILP 2: (AHB62359.1) insulin related peptide 2 neuropeptide precursor [Platynereis dumerilii]; HsIns: (AAA59172.1) insulin [Homo sapiens]. Putative monobasic/dibasic cleavage sites are indicated by blue blocks. Amino acid residues corresponding to signal peptide. The A chain, B chain and C chain are indicated by a purple, green, blue and orange background color, respectively. Disulfide bonds are indicated by black lines connecting the two corresponding cysteine residues.
Fig 7.
Multiple sequence alignment of BnVP-like with vasopressin and oxytoxin homologs, and VP-type peptide precursor from deuterostome and protostome taxa.
SpOxy: oxytocin-neurophysin 1 [Strongylocentrotus purpuratus] (XP_003724490.1); HsOxy:) oxytocin-neurophysin 1 preproprotein [Homo sapiens] (NP_000906.1); HsVas: (P_000481.2) vasopressin-neurophysin 2-copeptin preproprotein [Homo sapiens]; RnVas: (CAA25795.2) vasopressin precursor [Rattus norvegicus]; MmVas: (AAC42027.1) neurophysin II/vasopressin [Mus musculus]; DrOxy: (NP_840076.1) oxytocin-neurophysin 1 precursor [Danio rerio]; DrVas: NP_840078.1| vasopressin-neurophysin 2-copeptin precursor [Danio rerio]; GgVas: (NP_990516.1) vasotocin-neurophysin VT precursor [Gallus gallus]; XtVaso: XP_002936404.1| PREDICTED: vasopressin-neurophysin 2-copeptin [Xenopus (Silurana) tropicalis]; EfAnn: (CAD20057.2) annetocin precursor [Eisenia fetida]; PdVas: (ABR68852.1) vasotocin-neurophysin [Platynereis dumerilii]; DpVas: (EFX71881.1) putative vasopressin-like neuropeptide preprohormone [Daphnia pulex]; TcVas: (EFA09262.1) arginine vasopressin-like peptide [Tribolium castaneum]; OvCep: (BAC84978.1) cephalotocin precursor [Octopus vulgaris]; OvOct: (BAC82435.1) octopressin [Octopus vulgaris]; (AAB35220.1) conopressin [Lymnaea stagnalis]; CgCono: (XP_011434624.1) PREDICTED: conopressin/neurophysin-like [Crassostrea gigas]. Disulfide bridges are indicated by a bolded curve. Two amino acids inserted in BnVP-like are indicated by the black arrow. The BnVP-like sequence is underlined in red. Note the hydrophobic region near the N-terminus of the putative carrier peptide neurophysin is completely absent in BnVP-like.