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Abigail Roldán-Salgado, Celidee Sánchez-Barreto, Paul Gaytán
Since Green Fluorescent Protein (GFP) was first successfully expressed in heterologous systems in 1994, many genes encoding other natural autofluorescent proteins (AFPs) have been cloned and subsequently modified by protein engineering to improve their physicochemical properties. Throughout this twenty-two-year period, glycine 67 (Gly67) has been regarded as the only amino acid in the entire protein family that is essential for the formation of the different reported chromophores. In this work, we demonstrate that a synthetic gene encoding LanFP10-A, a natural protein encoded in the genome of the lancelet Branchiostoma floridae containing the G67A mutation, produces a heterologous, functional yellow fluorescent protein when expressed in E...
November 5, 2016: Gene
Jia-Xing Yue, Iryna Kozmikova, Hiroki Ono, Carlos W Nossa, Zbynek Kozmik, Nicholas H Putnam, Jr-Kai Yu, Linda Z Holland
Cephalochordates, the sister group of vertebrates + tunicates, are evolving particularly slowly. Therefore, genome comparisons between two congeners of Branchiostoma revealed so many conserved noncoding elements (CNEs), that it was not clear how many are functional regulatory elements. To more effectively identify CNEs with potential regulatory functions, we compared noncoding sequences of genomes of the most phylogenetically distant cephalochordate genera, Asymmetron and Branchiostoma, which diverged approximately 120-160 million years ago...
2016: Genome Biology and Evolution
Jia-Xing Yue, Nicholas D Holland, Linda Z Holland, Dimitri D Deheyn
Green Fluorescent Protein (GFP) was originally found in cnidarians, and later in copepods and cephalochordates (amphioxus) (Branchiostoma spp). Here, we looked for GFP-encoding genes in Asymmetron, an early-diverged cephalochordate lineage, and found two such genes closely related to some of the Branchiostoma GFPs. Dim fluorescence was found throughout the body in adults of Asymmetron lucayanum, and, as in Branchiostoma floridae, was especially intense in the ripe ovaries. Spectra of the fluorescence were similar between Asymmetron and Branchiostoma...
2016: Scientific Reports
Jing Wang, Guang Li, Guang-Hui Qian, Jun-Hao Hua, Yi-Quan Wang
The Wnt/β-catenin signaling pathway plays a crucial role in the embryonic development of metazoans. Although the pathway has been studied extensively in many model animals, its function in amphioxus, the most primitive chordate, remains largely uncharacterized. To obtain basic data for functional analysis, we identified and isolated seven genes (Lrp5/6, Dvl, APC, CkIα, CkIδ, Gsk3β, and Gro) of the Wnt/β-catenin signaling pathway from the amphioxus (Branchiostoma floridae) genome. Phylogenetic analysis revealed that amphioxus had fewer members of each gene family than that found in vertebrates...
May 18, 2016: Dong Wu Xue Yan Jiu, Zoological Research
Myles G Garstang, Peter W Osborne, David E K Ferrier
BACKGROUND: The ParaHox genes play an integral role in the anterior-posterior (A-P) patterning of the nervous system and gut of most animals. The ParaHox cluster is an ideal system in which to study the evolution and regulation of developmental genes and gene clusters, as it displays similar regulatory phenomena to its sister cluster, the Hox cluster, but offers a much simpler system with only three genes. RESULTS: Using Ciona intestinalis transgenics, we isolated a regulatory element upstream of Branchiostoma floridae Gsx that drives expression within the central nervous system of Ciona embryos...
2016: BMC Evolutionary Biology
Dong-Dong Cao, Xin Liao, Wang Cheng, Yong-Liang Jiang, Wen-Jie Wang, Qiong Li, Jun-Yuan Chen, Yuxing Chen, Cong-Zhao Zhou
Discovery of variable lymphocyte receptors (VLRs) in agnathans (jawless fish) has brought the origin of adaptive immunity system (AIS) forward to 500 million years ago accompanying with the emergence of vertebrates. Previous findings indicated that amphioxus, a representative model organism of chordate, also possesses some homologs of the basic components of TCR/BCR-based AIS, but it remains unknown if there exist any components of VLR-based AIS in amphioxus. Bioinformatics analyses revealed the amphioxus Branchiostoma floridae encodes a group of putative VLR-like proteins...
2016: Scientific Reports
Anjalika Pathirana, Mingyue Diao, Shibo Huang, Lingling Zuo, Yujun Liang
In fish, a series of maternal derived immune components have been identified in their eggs or embryos at very early stages, which are proposed to provide protections to themselves against pathogenic attacks from hostile environment. The phenomenon of maternal immunity has been also recorded in several invertebrate species, however, so far, very limited information about the maternal immune molecules are available. In this study, it was demonstrated maternal alpha2 macroglobulin (A2m) protein, an important innate immune factor, exists in the fertilized eggs of amphioxus Branchiostoma japonicum, an invertebrate chordate...
March 2016: Fish & Shellfish Immunology
Xiaojun Song, Juan Du, Wei Zhu, Ping Jin, Fei Ma
The ASPP (apoptosis-stimulating protein of p53) protein family plays very key roles in apoptosis regulation, in both p53-dependent and p53-independent pathways. However, the ASPP homologous gene has not been identified in amphioxus to date. Here, we identified and characterized an ASPP gene from Branchiostoma belcheri (designed as AmphiASPP) and extensively studied its evolution and roles involved in innate immunity. The results showed that the amphioxus genome has an ASPP homolog gene with an ORF of 3285 bp, encoding 1094 amino acids which contains ANK repeats and SK3 domain...
February 2016: Fish & Shellfish Immunology
Haohan Liu, Miaomiao Lei, Xiaoyuan Du, Pengfei Cui, Shicui Zhang
The emergence of multi-drug resistant (MDR) microbes leads to urgent demands for novel antibiotics exploration. We demonstrated a cDNA from amphioxus Branchiostoma japonicum, designated Bjamp1, encoded a protein with features typical of antimicrobial peptides (AMPs), which is not homologous to any AMPs currently discovered. It was found that Bjamp1 was expressed in distinct tissues, and its expression was remarkably up-regulated following challenge with LPS and LTA. Moreover, the synthesized putative mature AMP, mBjAMP1, underwent a coil-to-helix transition in the presence of TFE or SDS, agreeing well with the expectation that BjAMP1 was a potential AMP...
2015: Scientific Reports
Toshio Sekiguchi, Kenji Kuwasako, Michio Ogasawara, Hiroki Takahashi, Shin Matsubara, Tomohiro Osugi, Ikunobu Muramatsu, Yuichi Sasayama, Nobuo Suzuki, Honoo Satake
The calcitonin (CT)/CT gene-related peptide (CGRP) family is conserved in vertebrates. The activities of this peptide family are regulated by a combination of two receptors, namely the calcitonin receptor (CTR) and the CTR-like receptor (CLR), and three receptor activity-modifying proteins (RAMPs). Furthermore, RAMPs act as escort proteins by translocating CLR to the cell membrane. Recently, CT/CGRP family peptides have been identified or inferred in several invertebrates. However, the molecular characteristics and relevant functions of the CTR/CLR and RAMPs in invertebrates remain unclear...
January 29, 2016: Journal of Biological Chemistry
Takayuki Onai, Toshihiro Aramaki, Hidehiko Inomata, Tamami Hirai, Shigeru Kuratani
INTRODUCTION: Somites, blocks of mesoderm tissue located on either side of the neural tube in the developing vertebrate embryo, are derived from mesenchymal cells in the presomitic mesoderm (PSM) and are a defining characteristic of vertebrates. In vertebrates, the somite segmental boundary is determined by Notch signalling and the antagonistic relationship of the downstream targets of Notch, Lfng, and Delta1 in the anterior PSM. The presence of somites in the basal chordate amphioxus (Branchiostoma floridae) indicates that the last common ancestor of chordates also had somites...
2015: Zoological Letters
Shaochun Yuan, Tingting Zheng, Peiyi Li, Rirong Yang, Jie Ruan, Shengfeng Huang, Zhenxin Wu, Anlong Xu
The IFN regulatory factor (IRF) family encodes transcription factors that play important roles in immune defense, stress response, reproduction, development, and carcinogenesis. Although the origin of the IRF family has been dated back to multicellular organisms, invertebrate IRFs differ from vertebrate IRFs in genomic structure and gene synteny, and little is known about their functions. Through comparison of multiple amphioxus genomes, in this study we suggested that amphioxus contains nine IRF members, whose orthologs are supposed to be shared among three amphioxus species...
December 15, 2015: Journal of Immunology: Official Journal of the American Association of Immunologists
Wen-Jie Wang, Wang Cheng, Ming Luo, Qingyu Yan, Hong-Mei Yu, Qiong Li, Dong-Dong Cao, Shengfeng Huang, Anlong Xu, Roy A Mariuzza, Yuxing Chen, Cong-Zhao Zhou
Peptidoglycan recognition proteins (PGRPs), which have been identified in most animals, are pattern recognition molecules that involve antimicrobial defense. Resulting from extraordinary expansion of innate immune genes, the amphioxus encodes many PGRPs of diverse functions. For instance, three isoforms of PGRP encoded by Branchiostoma belcheri tsingtauense, termed BbtPGRP1~3, are fused with a chitin binding domain (CBD) at the N-terminus. Here we report the 2.7 Å crystal structure of BbtPGRP3, revealing an overall structure of an N-terminal hevein-like CBD followed by a catalytic PGRP domain...
2015: PloS One
Shousheng Liu, Yuanyuan Liu, Shuangshuang Yang, Youhua Huang, Qiwei Qin, Shicui Zhang
RIG-I-like (where RIG-I is retinoic acid inducible gene I) receptor LGP2 (where LGP2 is laboratory of genetics and physiology) is an important intracellular receptor that recognizes viral RNAs in innate immunity, but its origin and evolution remains unknown. Here we clearly demonstrate the presence of a RIG-I-like receptor, BjLGP2, in the basal chordate amphioxus. It is predominantly expressed in the hepatic caecum and hindgut, and is upregulated following challenge with poly(I:C). BjLGP2 is distributed in the cytoplasm of both grouper spleen and flounder gill (FG) cells, and the recombinant BjLGP2 interacts with poly(I:C)...
December 2015: European Journal of Immunology
Radhey S Gupta
Members of the phylum Chordata and the subphylum Vertebrata are presently distinguished solely on the basis of morphological characteristics. The relationship of the vertebrates to the two non-vertebrate chordate subphyla is also a subject of debate. Analyses of protein sequences have identified multiple conserved signature indels (CSIs) that are specific for Chordata or for Vertebrata. Five CSIs in 4 important proteins are specific for the Vertebrata, whereas two other CSIs are uniquely found in all sequenced chordate species including Ciona intestinalis and Oikapleura dioica (Tunicates) as well as Branchiostoma floridae (Cephalochordates)...
January 2016: Molecular Phylogenetics and Evolution
Huihui Wang, Baobao Liu, Hongyan Li, Shicui Zhang
Polyamine oxidases (PAOs) have been identified in a wide variety of animals, as well as in fungi and plant. Generally, plant PAOs oxidize spermine (Spm), spermidine (Spd) and their acetylated derivatives, N(1)-acetylspermine (N(1)-Aspm) and N(1)-acetylspermidine (N(1)-Aspd), while yeast PAOs oxidize Spm, N(1)-Aspm and N(1)-Aspd, but not Spd. By contrast, two different enzymes, namely spermine oxidase (SMO) and acetylpolyamine oxidase (APAO), specifically catalyze the oxidation of Spm and N(1)-Aspm/N(1)-Aspd, respectively...
January 10, 2016: Gene
Z B Li, Y S Huang, J B Shangguan, Y F Ning, Y Yi, G Dai
Branchiostoma belcheri Gray is a second-class, nationally protected protochordate in China. We developed 10 novel polymorphic sites in B. belcheri, which were examined using a population of 30 wild individuals from Xiamen, China. The polymorphism information content ranged between 0.141 and 0.681, and the number of alleles varied from 2 to 5. The expected and observed heterozygosities varied between 0.1528 and 0.6920, and between 0.1429 and 0.5000, respectively. These novel microsatellite markers will facilitate the genetic analysis and protection of wild B...
2015: Genetics and Molecular Research: GMR
Denghua Yin, Wenjuan Li, Meili Fu, Liming Chen, Fei Ma, Ping Jin
Transforming growth factor-β activated kinase-1 (TAK1) is an essential regulator in toll-like receptor (TLR), tumor necrosis factor (TNF) and interleukin-1 (IL-1) signaling pathways, and plays very important roles in animal innate immunity. TAK1-binding protein, TAB1, can specifically regulate the activation of TAK1. However, the TAB1 gene in amphioxus has not yet been identified to date. In this study, we identified and characterized a TAB1 gene from Branchiostoma belcheri (designed as AmphiTAB1). Our results showed that the full-length cDNA of AmphiTAB1 is 2281bp long with an open reading frame (ORF) of 1659bp that encodes a predicted protein of 553 amino acids containing a typical PP2Cc domain...
January 10, 2016: Gene
Seongsik Yun, Michael Furlong, Mikang Sim, Minah Cho, Sumi Park, Eun Bee Cho, Arfaxad Reyes-Alcaraz, Jong-Ik Hwang, Jaebum Kim, Jae Young Seong
In humans, numerous genes encode neuropeptides that comprise a superfamily of more than 70 genes in approximately 30 families and act mainly through rhodopsin-like G protein-coupled receptors (GPCRs). Two rounds of whole-genome duplication (2R WGD) during early vertebrate evolution greatly contributed to proliferation within gene families; however, the mechanisms underlying the initial emergence and diversification of these gene families before 2R WGD are largely unknown. In this study, we analyzed 25 vertebrate rhodopsin-like neuropeptide GPCR families and their cognate peptides using phylogeny, synteny, and localization of these genes on reconstructed vertebrate ancestral chromosomes (VACs)...
November 2015: Molecular Biology and Evolution
Jian Peng, Xin Tao, Rui Li, Jingru Hu, Jie Ruan, Ruihua Wang, Manyi Yang, Rirong Yang, Xiangru Dong, Shangwu Chen, Anlong Xu, Shaochun Yuan
Studies have shown that the basal chordate amphioxus possesses an extraordinarily complex TLR system, including 39 TLRs and at least 40 Toll/IL-1R homologous region (TIR) adaptors. Besides homologs to MyD88 and TIR domain-containing adaptor molecule (TICAM), most amphioxus TIR adaptors exhibit domain architectures that are not observed in other species. To reveal how these novel TIR adaptors function in amphioxus Branchiostoma belcheri tsingtauense (bbt), four representatives, bbtTIRA, bbtTIRB, bbtTIRC, and bbtTIRD, were selected for functional analyses...
October 1, 2015: Journal of Immunology: Official Journal of the American Association of Immunologists
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