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Molecular Cloning of MSRG-11 Gene Related to Apoptosis of Mouse Spermatogenic Cells

Acta Biochimica et Biophysica Sinica 2005年第3期37卷 159-166页

作者： Yun DENG Dong-Song NIE Jian WANG Xiao-Jun TAN Zhao-Yan NIE Hong-Mei YANG Liang-Sha HU Guang-Xiu LU Human Reproductive and Stem Cell Engineering Institute,Central South University,Changsha 410078,China Human Reproductive and Stem Cell Engineering Institute Central South UniversityChangsha 410078 China

Beginning with a new contig of the expressed sequence tags(Mm.63892)obtained bycomparing testis libraries with other tissue and cell line libraries using the digital differential display program,we cloned a new gene which is related to the apoptosis of mouse spermatogenic cells using the Genscanprogram and polymerase chain reaction(PCR)*** sequence data have been submitted to theGenBank database under accession number *** full cDNA length is 1074 bp,and the gene with7 exons and 6 introns is located in mouse chromosome 1 *** protein is recognized as a new member ofcahnodulin(CaM)binding protein family because the sequence contains three short calmodulin-binding motifscontaining conserved Ile and Gln residues(iq motif)and is considered to play a critical role in interactions ofiq motif-containing proteins with CaM *** putative protein encoded by this gene has 192 aminoacid residues with a theoretical molecular mass of 23.7 kDa and a calculated isoelectric point of *** shares no significant homology with any known protein in ***-PCR and Northern blotanalyses revealed that 1.3 kb MSRG-11 transcript was strongly expressed in adult mouse testis but weaklyexpressed in the spleen and *** MSRG-11 gene was expressed at various levels,faintly at twoweeks postpartum and strongly from three weeks postpartum in adult *** green fluorescence pro-duced by pEGFP-C2/MSRG-11 was detected in the cytoplasm of COS7 cells 24 h ***3.1(-)/MSRG-11 plasmid was constructed and introduced into COS7 cells using Lipofectamine 2000transfection reagent(Invitrogen,Carlsbad,USA).MSRG-11 can accelerate COS7 cell apoptosis,whichsuggests that this gene may play an important role in the development of mouse testes and is a candidate geneof testis-specific *** on these observations,it was considered that we cloned a new gene whichprobably accelerates spermatogenetic cell apoptosis in mouse.

关键词： MSRG-11 gene testis-specific expression spermatogenetic cell apoptosis iq motif

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Isolation and characterization of Xenopus laevis homologs of the mouse inv gene and functional analysis of the conserved calmodulin binding sites

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Cell Research 2006年第4期16卷 337-346页

作者： YukutoYasuhiko KoichiroShiokawa Toshio Mochizuki Makoto Asashima Takahiko Yokoyama Department of Biological Sciences Graduate School of ScienceUniversity of Tokyo7-3-1 HongoBunkyo-kuToyo113-0033Japan Department of Medicine Kidney CenterTokyo Women's Medical UniversitySchool of MedicineJapan Department of Life Science University of Tokyo3-8-1 Komaba.Meguro-kuTokyo 158-8902.Japan Department of Anatomy and Developmental Biology Tokyo Women's Medical College8-1 Kawada-choShinjuku-kuTokyo 162-8666Japan

The homozygous inv(inversion of embryonic turning)mouse mutant shows situs inversus and polycystic kidney disease,both of which result from the lack of the inv ***,we suggested that inv may be important for the ieft-rightaxis formation,not only in mice but also in Xenopus,and that calmodulin regulates this inv protein ***,weisolated and characterized two Xenopus laevis homologs(Xinv-1 and Xinv-2)of the mouse inv gene,and performedfunctional analysis of the conserved iq motifs that interact with ***-1 expresses early in development in thesame manner as mouse inv ***,a full-length Xenopus inv mRNA did not randomize cardiac orientationwhen injected into Xenopus embryos,which is different from mouse inv *** to mouse inv mRNA,Xenopusinv mRNA with mutated iq randomized cardiac *** present study indicates that calmodulin binding sites(iq motifs)are crucial in controlling the biological activity of both mouse and Xenopus inv *** mouseand Xenopus inv genes have a quite similar structure,the interaction with calmodulin and iq motifs ofXenopus inv andmouse inv proteins may regulate their function in different ways.

关键词： Left-right asymmetry inv calmodulin ankyrin iq motif Xenopus calcium

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拟南芥辅助分子伴侣突变体fes1a bag6以及fes1a hop1 hop2的研究

拟南芥辅助分子伴侣突变体fes1a bag6以及fes1a hop1 hop2的研究

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作者：侯燕飞山东师范大学

学位级别：硕士

HSP70s在植物的热胁迫中发挥着重要的作用。HSP70有许多辅助分子伴侣,其中BAG6和Fes1A作为HSP70的结合蛋白,即辅助分子伴侣,对拟南芥耐热性有着至关重要的作用。目前,对植物辅助分子伴侣之间如何相互作用知之甚少。本研究中,我们首先确... 详细信息

HSP70s在植物的热胁迫中发挥着重要的作用。HSP70有许多辅助分子伴侣,其中BAG6和Fes1A作为HSP70的结合蛋白,即辅助分子伴侣,对拟南芥耐热性有着至关重要的作用。目前,对植物辅助分子伴侣之间如何相互作用知之甚少。本研究中,我们首先确定BAG6的亚细胞定位。根据BAG6-GFP亚细胞荧光信号,以及纯化的细胞核和液泡组分western-blot分析,揭示BAG6蛋白在细胞核定位表达,而并非文献报道的液泡定位蛋白。通过对fes1a bag6双突变体的耐热性变化分析,我们发现,与fes1a相比,fes1a bag6双突变体的耐热性明显增加,但钙调蛋白抑制剂降低了fes1a bag6双突变体耐热性。实验还发现,BAG6中iq motif对于BAG6的功能至关重要。由于BAG6位于细胞核中,提示信号转导可能涉及核钙信号。此外,实验还发现BAG6对fes1a的调控与TBP对fes1a的调控归属不同的信号通路。Hop为HSP70的另一辅助分子伴侣。本实验室前期研究发现hop单突变体对盐不敏感,但hop1 hop2双突变体呈现盐敏感表型;对HSP70的另一辅助分子伴侣Fes1A而言,仅在萌发期且超过125 m M Na Cl胁迫下,fes1a呈现盐敏感表型。在本实验中发现,与hop1 hop2双突变体或fes1a相比,fes1a hop1 hop2三突变体对盐更敏感,在75 m M Na Cl胁迫时即出现明显的盐敏感表型。然而,fes1a hop1 hop2三突变体,hop1 hop2双突变体和fes1a在MS培养基上均正常生长,长势与WT野生型一致。这些结果提示HSP70的辅助分子伴侣Fes1A和Hop可能协同调控拟南芥的耐盐性。本课题主要创新点:1、iq motif对于BAG6功能的有效性至关重要;2、钙调蛋白参与信号转导;3、BAG6定位于细胞核中;4、BAG6与TBP对fes1a的调控归属不同的信号通路;5、Fes1A和Hop协同调控拟南芥的耐盐性。

关键词： Fes1A BAG6 抑制子钙调蛋白 iq motif TBP Hop 盐敏感性

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