Cloning and function analysis of BAG family genes in wheat
Shiming Ge A , Zhen Kang A , Ying Li A , Fuzhen Zhang A , Yinzhu Shen A , Rongchao Ge A B C and Zhanjing Huang A B CA College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei 050024, China.
B Corresponding authors. Emails: huangzhanjing@hebtu.edu.cn; grcgp@sina.com
C These authors contributed equally to this work.
Functional Plant Biology 43(5) 393-402 https://doi.org/10.1071/FP15317
Submitted: 6 October 2015 Accepted: 13 January 2016 Published: 23 February 2016
Abstract
By analysing the cDNA microarray of the salt tolerant mutant of wheat RH8706–49 under salinity stress, our results showed an expressed sequence tag fragment and acquired an unknown gene (designated as TaBAG) with a BAG conserved domain through electronic cloning and RT–PCR technology. The gene was registered into GenBank (No. FJ599765). After homologous alignment analysis, electronic cloning, and amplifying with RT–PCR, the other gene with a BAG conserved domain, TaBAG2, was obtained and registered into GenBank (No. GU471210). Quantitative PCR analysis demonstrated that TaBAG2 expression was induced by saline and heat stress. TaBAG gene expression under salinity stress increased remarkably but showed an insignificant response to heat stress. The adversity stress detection results showed that Arabidopsis overexpressing TaBAG and TaBAG2 exhibited an obvious salt tolerance increase. Under heat stress, Arabidopsis overexpressing TaBAG2 showed increased heat tolerance; however, the heat tolerance of Arabidopsis overexpressing TaBAG did not vary significantly under heat stress. Subcellular localisation results demonstrated that TaBAGs were mainly located in the cytoplasm and the cell nucleus. We applied fluorescence complementation and yeast two-hybrid technique to prove that TaBAG2 can obviously bond with TaHsp70 and TaCaMs. After the respective mutation of aspartic acid (D) and arginine (R) at high conservation in BAG domain of TaBAG2, the bonding interaction between TaBAG2 and TaHsp70 disappeared, indicating that the two amino acids were the key loci for the interaction between TaBAG2 and TaHsp70. Heat tolerance detection results demonstrated that the heat tolerance of Arabidopsis overexpressing and cotransfected with TaBAG2 and TaHsp70 was much higher than that of Arabidopsis overexpressing TaBAG2 and Arabidopsis overexpressing TaHSP70. This finding implies that the synergistic use of TaBAG2 and TaHSP70 can improve heat tolerance of plants.
Additional keywords: BAG family, CaM, heat tolerance, signaling pathway, signalling pathway, TaHsp70, Triticum aestivum.
References
Antoku K, Maser RS, Scully WJ, Delach SM, Johnson DE (2001) Isolation of Bcl-2 binding proteins that exhibit homology with BAG-1 and suppressor of death domains protein. Biochemical and Biophysical Research Communications 286, 1003–1010.| Isolation of Bcl-2 binding proteins that exhibit homology with BAG-1 and suppressor of death domains protein.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXmtlejtbc%3D&md5=0487e5cfc368eca36998d5d15e7cc9bdCAS | 11527400PubMed |
Bannai H, Tamada Y, Maruyama O, Nakai K, Miyano S (2002) Extensive feature detection of N-terminal protein sorting signals. Bioinformatics 18, 298–305.
| Extensive feature detection of N-terminal protein sorting signals.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XitFensb4%3D&md5=3ae8daea14142b760fa19b6ee127b9a5CAS | 11847077PubMed |
Bardelli A, Longati P, Albero D, Goruppi S, Schneider C, Ponzetto C, Comoglio PM (1996) HGF receptor associates with the anti-apoptotic protein BAG-1 and prevents cell death. EMBO Journal 15, 6205–6212.
Boyer JS (1982) Plant productivity and environment. Science 218, 443–448.
| Plant productivity and environment.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3cvjvVahuw%3D%3D&md5=99ce43610750779c6119ce3239a6fedfCAS | 17808529PubMed |
Cheney RE, Mooseker MS (1992) Unconventional myosins. Current Opinion in Cell Biology 4, 27–35.
| Unconventional myosins.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK38Xhs1ynsLs%3D&md5=facc7229f0ea5567eaadeaedd6654b9bCAS | 1558751PubMed |
Doong H, Vrailas A, Kohn EC (2002) What’s in the ‘BAG’? – A functional domain analysis of the BAG-family proteins. Cancer Letters 188, 25–32.
| What’s in the ‘BAG’? – A functional domain analysis of the BAG-family proteins.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38Xot1Wqt7k%3D&md5=4dfa5ec18656a9263e4b516152fe70a3CAS | 12406544PubMed |
Doukhanina EV, Chen S, van der Zalm E, Godzik A, Reed J, Dickman MB (2006) Identification and functional characterization of the BAG protein family in Arabidopsis thaliana. Journal of Biological Chemistry 13, 84–95.
Emanuelsson O, Nielsen H, Brunak S, Heijne G (2000) Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. Journal of Molecular Biology 300, 1005–1016.
| Predicting subcellular localization of proteins based on their N-terminal amino acid sequence.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXks1OntrY%3D&md5=87c3600bc21b81284384d60e9bb9610cCAS | 10891285PubMed |
Froesch BA, Takayama S, Reed JC (1998) BAG-1L protein enhances androgen receptor function. The Journal of Biological Chemistry 273, 11660–11666.
| BAG-1L protein enhances androgen receptor function.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXjt1Wrt7s%3D&md5=c8c4e0ca5d357d6e315ca7cced002cb1CAS | 9565586PubMed |
Hu LZ, Chen JT, Guo JJ, Zhao YF, Zhu LY, Huang YQ (2013) Functional divergence and evolutionary dynamics of BAG gene family in maize (Zea mays). International Journal of Agriculture and Biology 2, 200–206.
Kang CH, Jung WY, Kang YH, Kim JY, Kim DG, Jeong JC, Baek DW, Jin JB, Lee JY, Kim MO, Chung WS, Mengiste T, Koiwa H, Kwak SS, Bahk JD, Lee SY, Nam JS, Yun DJ, Cho MJ (2006) AtBAG6, a novel calmodulin-binding protein, induces programmed cell death in yeast and plants. Cell Death and Differentiation 13, 84–95.
| AtBAG6, a novel calmodulin-binding protein, induces programmed cell death in yeast and plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXht1yktbrL&md5=8bc64e90a20a8fdc26f7c1d4b7c6c09bCAS | 16003391PubMed |
Kim MC, Panstruga R, Elliott C, Müller J, Devoto A, Yoon HW, Park HC, Cho MJ, Schulze-Lefert P (2002) Calmodulin interacts with MLO protein toregulate defence against mildew in barley. Nature 416, 447–451.
| Calmodulin interacts with MLO protein toregulate defence against mildew in barley.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XislamsbY%3D&md5=082584c89b3fed9bd322be0c6efb297eCAS | 11919636PubMed |
Kobayashi M, Katoh H, Takayanagi T, Suzuki S (2010) Characterization of hermotolerance-related genes in grapevine (Vitis vinifera). Journal of Plant Physiology 167, 812–819.
| Characterization of hermotolerance-related genes in grapevine (Vitis vinifera).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXosFChtLc%3D&md5=6d9b8e7c84443e475b2a68d8fc1be24fCAS | 20096476PubMed |
Krasensky J, Jonak C (2012) Drought, salt, and temperature stress-induced metabolic rearrangements and regulatory networks. Journal of Experimental Botany 63, 1593–1608.
| Drought, salt, and temperature stress-induced metabolic rearrangements and regulatory networks.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XjtlKhs7w%3D&md5=e7346d63aa4ee511534e53ac1d5adf0bCAS | 22291134PubMed |
Kullmann M, Schneikert J, Moll J, Heck S, Zeiner M, Gehring U, Cato AC (1998) RAP46 is a negative regulator of glucocorticoid receptor action and hormone-induced apoptosis. Journal of Biological Chemistry 273, 14620–14625.
| RAP46 is a negative regulator of glucocorticoid receptor action and hormone-induced apoptosis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXjs1OqsLk%3D&md5=de75b3204df5e6cc26d9f4f15fa4205cCAS | 9603979PubMed |
Li B, Liu HT, Mu RL (2002) Effects of calmodulin on DNA-binding activity of heat shock transcription factor in vitro. Chinese Science Bulletin 3, 255–258.
Liu R, Takayama S, Zheng Y, Froesch B, Chen G, Zhang X, Reed JC, Zhang X (1998) Interaction of BAG-1 with retinoic acid receptor and its inhibition of retinoic acid-induced apoptosis in cancer cells. Journal of Biological Chemistry 273, 16985–16992.
| Interaction of BAG-1 with retinoic acid receptor and its inhibition of retinoic acid-induced apoptosis in cancer cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXkt12ntbo%3D&md5=9e1234c38bdbc806013fe475685f0a00CAS | 9642262PubMed |
Liu HT, Li B, Shang ZL, Li XZ, Mu RL, Sun DY, Zhou RG (2003) Calmodulin is involved in heat shock signal transduction in wheat. Plant Physiology 132, 1186–1195.
| Calmodulin is involved in heat shock signal transduction in wheat.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXlsFGht7k%3D&md5=47d0d531cc7e3502af6da7c88165b7c5CAS | 12857801PubMed |
Naishiro Y, Adachi M, Okuda H, Yawata A, Mitaka T, Takayama S, Reed JC, Hinoda Y, Imai K (1999) BAG-1 accelerates cell motility of human gastric cancer cells. Oncogene 18, 3244–3251.
| BAG-1 accelerates cell motility of human gastric cancer cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXjvVeqs78%3D&md5=5ad4a22824ad754e398918c853ef7cd8CAS | 10359530PubMed |
Nollen EAA, Brunsting JF, Song J, Kampinga HH, Morimoto RI (2000) Bag-1 functions in vivo as a negative regulator of Hsp70 chaperone activity. Molecular and Cellular Biology 20, 1083–1088.
| Bag-1 functions in vivo as a negative regulator of Hsp70 chaperone activity.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXmt1Smuw%3D%3D&md5=a62600b55f724893ff1e5117a17f1697CAS |
Schneikert J, Hübner S, Martin E, Cato AC (1999) A nuclear action of the eukaryotic cochaperone RAP46 in downregulation of glucocorticoid receptor activity. Journal of Cell Biology 146, 929–940.
| A nuclear action of the eukaryotic cochaperone RAP46 in downregulation of glucocorticoid receptor activity.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXlvVKjsbo%3D&md5=938e46bf8ef481e1182feb518640be88CAS | 10477749PubMed |
Sondermann H, Scheufler C, Schneider C, Hohfeld J, Hartl FU, Moarefi I (2001) Structure of a Bag/Hsc70 complex: convergent functional evolution of Hsp70 nucleotide exchange factors. Science 291, 1553–1557.
| Structure of a Bag/Hsc70 complex: convergent functional evolution of Hsp70 nucleotide exchange factors.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXhsVeqt7s%3D&md5=8ced23bdf62236276b38f1906c63ba23CAS | 11222862PubMed |
Suzuki M, Youle RJ, Tjandra N (2000) Structure of Bax: coregulation of dimer formation and intracellular localization. Cell 103, 645–654.
| Structure of Bax: coregulation of dimer formation and intracellular localization.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXot1Gnu7c%3D&md5=8565a67416924da30d34be866013840fCAS | 11106734PubMed |
Takayama S, Xie Z, Reed JC (1999) An evolutionarily conserved family of Hsp70/Hsc70 molecular chaperone regulators. Journal of Biological Chemistry 274, 781–786.
| An evolutionarily conserved family of Hsp70/Hsc70 molecular chaperone regulators.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXmtFGmtg%3D%3D&md5=26acb7aaa778f9e770f70b7a7f0e5dfbCAS | 9873016PubMed |
Townsend PA, Cutress RI, Sharp A, Brimmell M, Packham G (2003) BAG-1: a multifunctional regulator of cell growth and survival. Biochimica et Biophysica Acta 1603, 83–98.
Wang HG, Takayama S, Rapp UR, Reed JC (1996) Bcl-2 interacting protein, BAG-1, binds to and activates the kinase Raf-1. Proceedings of the National Academy of Sciences of the United States of America 93, 7063–7068.
| Bcl-2 interacting protein, BAG-1, binds to and activates the kinase Raf-1.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28Xkt1Wht78%3D&md5=db8977e0d7ecbd9f9840a444f47ef960CAS | 8692945PubMed |
Zeiner M, Niyaz Y, Gehring U (1999) The hsp70-associating protein Hap46 binds to DNA and stimulates transcription. Proceedings of the National Academy of Sciences of the United States of America 96, 10194–10199.
| The hsp70-associating protein Hap46 binds to DNA and stimulates transcription.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXlvFegtbY%3D&md5=e20d11643b977701863352a3f7cbe1a8CAS | 10468585PubMed |
