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Proteomic characterization and evolutionary analyses of zona pellucida domain-containing proteins in the egg coat of the cephalochordate, Branchiostoma belcheri
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  • 作者:Qianghua Xu (1)
    Guang Li (2)
    Lixue Cao (3)
    Zhongjun Wang (1)
    Hua Ye (3)
    Xiaoyin Chen (2)
    Xi Yang (2)
    Yiquan Wang (2)
    Liangbiao Chen (3) (4)
  • 关键词:Amphioxus ; Zona pellucida protein ; Proteomics ; Molecular evolution ; Sperm ; egg interaction
  • 刊名:BMC Evolutionary Biology
  • 出版年:2012
  • 出版时间:December 2012
  • 年:2012
  • 卷:12
  • 期:1
  • 全文大小:1414KB
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  • 作者单位:Qianghua Xu (1)
    Guang Li (2)
    Lixue Cao (3)
    Zhongjun Wang (1)
    Hua Ye (3)
    Xiaoyin Chen (2)
    Xi Yang (2)
    Yiquan Wang (2)
    Liangbiao Chen (3) (4)

    1. Key Laboratory of Sustainable Exploitation of Oceanic Fisheries Resources, Ministry of Education, College of Marine Sciences, Shanghai Ocean University, Shanghai, People鈥檚 Republic of China
    2. School of Life Sciences, Xiamen University, Xiamen, People鈥檚 Republic of China
    3. State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, People鈥檚 Republic of China
    4. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Nan-Yi-Tiao Road #3, ZhongGuanCun, Beijing, 100190, People鈥檚 Republic of China
文摘
Background Zona pellucida domain-containing proteins (ZP proteins) have been identified as the principle constituents of the egg coat (EC) of diverse metazoan taxa, including jawed vertebrates, urochordates and molluscs that span hundreds of millions of years of evolutionary divergence. Although ZP proteins generally contain the zona pellucida (ZP) structural modules to fulfill sperm recognition and EC polymerization functions during fertilization, the primary sequences of the ZP proteins from the above-mentioned animal classes are drastically different, which makes it difficult to assess the evolutionary relationships of ZP proteins. To understand the origin of vertebrate ZP proteins, we characterized the egg coat components of Branchiostoma belcheri, an invertebrate species that belongs to the chordate subphylum Cephalochordata. Results Five ZP proteins (BbZP1-5) were identified by mass spectrometry analyses using the egg coat extracts from both unfertilized and fertilized eggs. In addition to the C-terminal ZP module in each of the BbZPs, the majority contain a low-density lipoprotein receptor domain and a von Willebrand factor type A (vWFA) domain, but none possess an EGF-like domain that is frequently observed in the ZP proteins of urochordates. Fluorescence in situ hybridization and immuno-histochemical analyses of B. belcheri ovaries showed that the five BbZPs are synthesized predominantly in developing eggs and deposited around the extracellular space of the egg, which indicates that they are bona fide egg coat ZP proteins. BbZP1, BbZP3 and BbZP4 are significantly more abundant than BbZP2 and BbZP5 in terms of gene expression levels and the amount of mature proteins present on the egg coats. The major ZP proteins showed high polymorphism because multiple variants are present with different molecular weights. Sequence comparison and phylogenetic analysis between the ZP proteins from cephalochordates, urochordates and vertebrates showed that BbZP1-5 form a monophyletic group and share no significant sequence similarities with the ZP proteins of urochordates and the ZP3 subtype of jawed vertebrates. By contrast, small regions of homology were identifiable between the BbZP and ZP proteins of the non-jawed vertebrate, the sea lamprey Petromyzon marinus. The lamprey ZP proteins were highly similar to the ZP1 and ZP2 subtypes of the jawed vertebrates, which suggests that the ZP proteins of basal chordates most likely shared a recent common ancestor with vertebrate ZP1/2 subtypes and lamprey ZP proteins. Conclusions The results document the spectra of zona pellucida domain-containing proteins of the egg coat of basal chordates. Particularly, the study provides solid evidence for an invertebrate origin of vertebrate ZP proteins and indicates that there are diverse domain architectures in ZP proteins of various metazoan groups.

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