priya kaur - Academia.edu (original) (raw)
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UERJ - Universidade do Estado do Rio de Janeiro / Rio de Janeiro State University
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Papers by priya kaur
PLOS Genetics
Gametogenesis requires coordinated signaling between germ cells and somatic cells. We previously ... more Gametogenesis requires coordinated signaling between germ cells and somatic cells. We previously showed that Gap junction (GJ)-mediated soma-germline communication is essential for fly spermatogenesis. Specifically, the GJ protein Innexin4/Zero population growth (Zpg) is necessary for somatic and germline stem cell maintenance and differentiation. It remains unknown how GJ-mediated signals regulate spermatogenesis or whether the function of these signals is restricted to the earliest stages of spermatogenesis. Here we carried out comprehensive structure/function analysis of Zpg using insights obtained from the protein structure of innexins to design mutations aimed at selectively perturbing different regulatory regions as well as the channel pore of Zpg. We identify the roles of various regulatory sites in Zpg in the assembly and maintenance of GJs at the plasma membrane. Moreover, mutations designed to selectively disrupt, based on size and charge, the passage of cargos through the...
Gap junction channels are intercellular transmembrane proteins that play an essential role in the... more Gap junction channels are intercellular transmembrane proteins that play an essential role in the development of tissues. They are a complex of proteins that can form homomeric or heteromeric channels between coupled cells to allow the passage of ions and small molecules. In Drosophila melanogaster, gap junctions are composed of innexins (Inx), which are structural homologues of vertebrate connexins. In Drosophila testes, Inx proteins contribute to spermatogenesis via bidirectional soma-germline communication. Inx2 on the somatic cells, which support the developing germline, and Zpg (Inx4) on the germline cells form a coupled heterotypic transmembrane channel in the fly testis. While the significance of these proteins is established, little is known about the nature of this intercellular communication. Our lab has previously demonstrated that flies lacking innexin expression have rudimentary gonads and impaired germline and somatic cell differentiation. The recently determined 3D protein structure of an innexin protein predicts that the N-terminus may be involved in regulating channel permeability. We are undertaking a detailed structure function analysis of innexin function in the context of the Drosophila testes using site-directed mutagenesis to generate mutations within the N-terminal and C-terminal domains that are predicted to disrupt functionally important residues. The effects on spermatogenesis are analysed using immunostainings and various functional assays. A 4 aminoacid truncation of the N-terminal results in a loss-of-function phenotype, and a deletion of the Cterminal disrupts the subcellular localization of the Zpg protein. This provides a mechanistic insight into the function of structurally important domains in innexins and further our understanding of germline stem cell regulation and maintenance during spermatogenesis.
PLOS Genetics
Gametogenesis requires coordinated signaling between germ cells and somatic cells. We previously ... more Gametogenesis requires coordinated signaling between germ cells and somatic cells. We previously showed that Gap junction (GJ)-mediated soma-germline communication is essential for fly spermatogenesis. Specifically, the GJ protein Innexin4/Zero population growth (Zpg) is necessary for somatic and germline stem cell maintenance and differentiation. It remains unknown how GJ-mediated signals regulate spermatogenesis or whether the function of these signals is restricted to the earliest stages of spermatogenesis. Here we carried out comprehensive structure/function analysis of Zpg using insights obtained from the protein structure of innexins to design mutations aimed at selectively perturbing different regulatory regions as well as the channel pore of Zpg. We identify the roles of various regulatory sites in Zpg in the assembly and maintenance of GJs at the plasma membrane. Moreover, mutations designed to selectively disrupt, based on size and charge, the passage of cargos through the...
Gap junction channels are intercellular transmembrane proteins that play an essential role in the... more Gap junction channels are intercellular transmembrane proteins that play an essential role in the development of tissues. They are a complex of proteins that can form homomeric or heteromeric channels between coupled cells to allow the passage of ions and small molecules. In Drosophila melanogaster, gap junctions are composed of innexins (Inx), which are structural homologues of vertebrate connexins. In Drosophila testes, Inx proteins contribute to spermatogenesis via bidirectional soma-germline communication. Inx2 on the somatic cells, which support the developing germline, and Zpg (Inx4) on the germline cells form a coupled heterotypic transmembrane channel in the fly testis. While the significance of these proteins is established, little is known about the nature of this intercellular communication. Our lab has previously demonstrated that flies lacking innexin expression have rudimentary gonads and impaired germline and somatic cell differentiation. The recently determined 3D protein structure of an innexin protein predicts that the N-terminus may be involved in regulating channel permeability. We are undertaking a detailed structure function analysis of innexin function in the context of the Drosophila testes using site-directed mutagenesis to generate mutations within the N-terminal and C-terminal domains that are predicted to disrupt functionally important residues. The effects on spermatogenesis are analysed using immunostainings and various functional assays. A 4 aminoacid truncation of the N-terminal results in a loss-of-function phenotype, and a deletion of the Cterminal disrupts the subcellular localization of the Zpg protein. This provides a mechanistic insight into the function of structurally important domains in innexins and further our understanding of germline stem cell regulation and maintenance during spermatogenesis.