Bioinformatic analysis of embryo development related small heat shock protein Hsp26 in <italic>Artemia</italic> species

doi:10.1007/s11515-012-1190-6

Front Biol

2012, Vol. 7

Issue (4) : 350-358 https://doi.org/10.1007/s11515-012-1190-6

RESEARCH ARTICLE

Bioinformatic analysis of embryo development related small heat shock protein Hsp26 in Artemia species

Jiaqing WANG¹, Lin HOU²(

), Zhenfeng HE³, Daizong Li⁴, Lijuan JIANG²

1. Science and Technology College, Shenyang Agricultural University, Fushun 113122, China; 2. College of Life Sciences, Liaoning Normal University, Dalian 116029, China; 3. State Key Laboratory of Supramolecular Structure and Materials, Jilin University, Changchun 130012, China; 4. College of Ocean, Agriculture University of Hebei, Qinhuangdao 066003, China

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Abstract

Artemia embryos can endure extreme temperature, long-term anoxia, desiccation and other wide variety of stressful conditions. How the embryos survive these stresses is a very interesting and unsolved subject. To solve this question we analyzed the nucleotide and deduced protein sequence for Hsp26, a molecular chaperone specific to Artemia embryo development. cDNAs of Hsp26 were sequenced from eight Artemia species and deduced Hsp26 amino acid sequences were analyzed. Computer-assisted analysis indicated that the 5′-untranslated region and all the 3 introns contain many putative cis-acting elements for Hsp26 gene expression during development, including heat shock elements (HSEs), Dfd, dl, CF2-II, Hb and AP-1 binding sites. Secondary structure of the Hsp26 3′-untranslated terminator contains the basic structure basis for transcriptional termination. Hsp26 shares sequence similarity with sHSPs (small heat shock protein) from other organisms. The physico-chemical properties of the deduced protein, such as theoretical molecular weight, protein extinction coefficient, isoelectric point and antigenic sites were also obtained. One seven-peptide nuclear localization signals (NLS) “PFRRRMM” was found, which suggested that the Hsp26 protein was hypothesized to be located inside the nucleus. The numbers of phosphorylation sites of serine, threonine and tyrosine and kinase specific phosphorylation sites are also located in Hsp26 protein sequence. These studies will help us achieve a better understanding of Hsp26 and broad implications for sHSPs function in crustacean embryo development.

Keywords bioinformatic analysis embryo development small heat shock protein Artemia species

Corresponding Author(s): HOU Lin,Email:houlin01@126.com

Issue Date: 01 August 2012

Cite this article:

Jiaqing WANG,Zhenfeng HE,Daizong Li, et al. Bioinformatic analysis of embryo development related small heat shock protein Hsp26 in Artemia species[J]. Front Biol, 2012, 7(4): 350-358.

URL:

https://academic.hep.com.cn/fib/EN/10.1007/s11515-012-1190-6
https://academic.hep.com.cn/fib/EN/Y2012/V7/I4/350

Tab.1 Ten districts of populations used in this study

Fig.1 Agarose gel electrophoresis analysis of total RNA of eight populations. Note: the abbreviation of species see Table. 1.

Fig.2 Nucleotide and deduced amino-acid sequence of the cDNA product for Hsp26 from the (YC). Note: The bold letters in boxes are the start codon (ATG) and the stop codon (TAA). The polyadenylation signal sequence (AATAAA) is shown in bold italic letters. The 3′-untranslated region (3′-UTR) used for secondary structure prediction are single underlined. The encoded protein contains two antigenic sites are shown in bold italic letters. The phosphorylation sites of serine, threonine and tyrosine are shaded gray and nuclear localization signal (NLS) is boxed.

Fig.3 Organization of Hsp26 exon and intron positions. Note: Schematic representation of Hsp26 genomic sequence and cDNA sequence were drawn to scale and aligned. E1–E4, exons; I1–I3, introns. Numbers above each schematic indicate Hsp26 exon and intron lengths.

Fig.4 Secondary structure of the Hsp26 terminator region calculated by RNAstructure 4.4 software.

Fig.5 Multiple sequence alignments of Hsp26 in ten populations. Note: The amino acid sequences were analyzed by CLUSTAL X (1.81) and the abbreviation of species see Table 1. Hsp26 domains based on the sequence of Hsp26 are indicated below the alignment, secondary structure elements based on the sequence of Hsp26 are above the alignment. Boxed sequences denoted the special motifs: glycine-enriched motif, arginine-enriched motif and threonine and serine-enhanced motif. Residue number is indicated on the right. –, no residue; *, identical residues;:, conserved substitution; ., semiconserved substitution.

Tab.2 Comparison of ten Hsp26 proteins in

Fig.6 Phylogenetic relationships between Hsp26 and the other 14 sHSP protein sequences. Note: One thousand bootstrap trials were run using the neighbor-joining algorithm by Mega 3.1 program. Square (?) indicates sHSPs from species. The species name and accession numbers of the used protein sequences are as follows: , ADD20465; , NP_523827; , XP_001663499; , XP_968760; , XP_001600020; , ACM24346; , AAZ14792; , BAF03558; , BAE94664; , ABC84494; , NP_037067; , NP_776715; , AAA18336; , AAA62175. The species sequences from GenBank are also as follows: UM, DQ310580; TK, DQ310579; AP, DQ310578; SFB, DQ310577; YC, DQ310576; GSL, AF031367.

1	Arrigo A P, Firdaus W J J, Mellier G, Moulin M, Paul C, Diaz-Iatoud C, Kretz-remy C (2005). Cytotoxic effects induced by oxidative stress in cultured mammalian cells and protection provided by Hsp27 expression. Methods , 35(2): 126–138 doi: 10.1016/j.ymeth.2004.08.003
2	Arrigo A P, Welch W J (1987). Characterization and purification of the small 28000-dalton marmmalian heat shock protein. J Biol Chem , 262(32): 15359–15369
3	Candido E P M (2002). The small heat shock proteins of the nematode Caenorhabditis elegans: structure, regulation and biology. In Small Stress Proteins : Arrigo A P, Muller W E G, (Eds.), Springer, Berlin, 61–79
4	Clegg J S (1994). Unusual response of Artemia franciscana embryos to prolonged anoxia. J Exp Zool , 270(3): 332–334 doi: 10.1002/jez.1402700312
5	Clegg J S, Jackson S A, Liang P, MacRae T H (1995). Nuclear-cytoplasmic translocations of protein p26 during aerobic-anoxic transitions in embryos of Artemia franciscana. Exp Cell Res , 219(1): 1–7 doi: 10.1006/excr.1995.1197
6	Coca M A, Almoguera C, Thomas T L, Jordano J (1996). Differential regulation of small heat-shock genes in plants: analysis of a water-stress-inducible and developmentally activated sunflower promoter. Plant Mol Biol , 31(4): 863–876 doi: 10.1007/BF00019473
7	Davidson S M, Loones M T, Duverger O, Morange M (2002). The developmental expression of small HSP, (Arrigo AP, Müller WEG, Eds.). Small Stress Proteins, Springer, Berlin, 103–128
8	Drinkwater L E, Crowe J H (1987). Regulation of embryonic diapause in Artemia: environmental and physiological signals. J Exp Zool , 241(3): 297–307 doi: 10.1002/jez.1402410304
9	Fan G C, Ren X, Qian J, Yuan Q, Nicolaou P, Wang Y, Jones K, Chu G, Kranias E G (2005). Novel cardioprotective role of a small heat-shock protein, Hsp20, against ischemia/reperfusion injury. Circulation , 111(14): 1792–1799 doi: 10.1161/01.CIR.0000160851.41872.C6
10	Gopal-Srivastava R, Cvekl A, Piatigorsky J (1998). Involvement of retinoic acid/retinoid receptors in the regulation of murine α B-crystallin/small heat shock protein gene expression in the lens. J Biol Chem , 273(28): 17954–17961 doi: 10.1074/jbc.273.28.17954
11	Guan J C, Jinn T L, Yeh C H, Feng S P, Chen Y M, Lin C Y (2004). Characterization of the genomic structures and selective expression profiles of nine class I small heat shock protein genes clustered on two chromosomes in rice (Oryza sativa L.). Plant Mol Biol , 56(5): 795–809 doi: 10.1007/s11103-004-5182-z
12	Guo W C, Chen A M (2008). Expression of heat shock protein gp96 in osteosarcoma and its clinical significance. Front Med China , 2(2): 200–203 doi: 10.1007/s11684-008-0038-6
13	Hand S C, Gnaiger E (1988). Anaerobic dormancy quantified in Artemia embryos: a calorimetric test of the control mechanism. Science , 239(4846): 1425–1427 doi: 10.1126/science.239.4846.1425
14	Haynes J I, Duncan M K, Piatigorsky J (1996). Spatial and temporal activity of the alpha B-crystallin/small heat shock protein gene promoter in transgenic mice. Dev Dyn , 207(1): 75–88 doi: 10.1002/(SICI)1097-0177(199609)207:1<75::AID-AJA8>3.0.CO;2-T
15	Ilagan J G, Cvekl A, Kantorow M, Piatigorsky J, Sax C M (1999). Regulation of α A-crystallin gene expression: lens specificity achieved through the differential placement of similar transcriptional control elements in mouse and chicken. J Biol Chem , 274(28): 19973–19978 doi: 10.1074/jbc.274.28.19973
16	Jackson S A, Clegg J S (1996). Ontology of low molecular weight stress protein p26 during early development of the brine shrimp, Artemia franciscana. Dev Growth Differ , 38(2): 153–160 doi: 10.1046/j.1440-169X.1996.t01-1-00004.x
17	Jakob U, Gaestel M, Engel K, Buchner J (1993). Small heat shock proteins are molecular chaperones. J Biol Chem , 268: 1517–1520
18	Jiang L J, Hou L, Zou X Y, Zhang R F, Wang J Q, Sun W J, Zhao X T, An J L (2007). Cloning and expression analysis of p26 gene in Artemia sinica. Acta Biochim Biophys Sin (Shanghai) , 39(5): 351–358 doi: 10.1111/j.1745-7270.2007.00287.x
19	Kato K, Goto S, Hasegawa K, Inaguma Y (1993). Coinduction of two low-molecular-weight stress proteins, alpha B crystallin and HSP28, by heat or arsenite stress in human glioma cells. J Biochem , 114: 640–647
20	Kato K, Goto S, Inaguma Y, Hasegawa K, Morishita R, Asano T (1994). Purification and characterization of a 20-kDa protein that is highly homologous to alpha B crystalline. J Biol Chem , 269: 15302– 15309
21	Kato K, Ito H, Inaguma Y (2002). Expression and phosphorylation of mammalian small heat shock proteins, In: Arrigo A P, Müller W E G (Eds.), Small Stress Proteins , Springer, Berlin, 127–150
22	Laksanalamai P, Robb F T (2004). Small heat shock proteins from extremophiles: a review. Extremophiles , 8(1): 1–11 doi: 10.1007/s00792-003-0362-3
23	Liang P, Amons R, Clegg J S, MacRae T H (1997). Molecular characterization of a small heat shock/α-crystallin protein in encysted Artemia embryos. J Biol Chem , 272(30): 19051–19058 doi: 10.1074/jbc.272.30.19051
24	Liang P, MacRae T H (1999). The synthesis of a small heat shock /α-crystallin protein in Artemia and its relationship to stress tolerance during development. Dev Biol , 207(2): 445–456 doi: 10.1006/dbio.1998.9138
25	Linder B, Jin Z, Freedman J H, Rubin C S (1996). Molecular characterization of a novel, developmentally regulated small embryonic chaperone from Caenorhabditis elegans. J Biol Chem , 271(47): 30158–30166 doi: 10.1074/jbc.271.47.30158
26	MacRae T H (2003). Molecular chaperones, stress resistance and development in Artemia franciscana. Semin Cell Dev Biol , 14(5): 251–258 doi: 10.1016/j.semcdb.2003.09.019
27	Maimbo M, Ohnishi K, Hikichi Y, Yoshioka H, Kiba A (2007). Induction of a small heat shock protein and its functional roles in Nicotiana plants in the defense response against Ralstonia solanacearum. Plant Physiol , 145(4): 1588–1599 doi: 10.1104/pp.107.105353
28	Marin R, Landry J, Tanguay R M (1996). Tissue-specific posttranslational modification of the small heat shock protein HSP27 in Drosophila. Exp Cell Res , 223(1): 1–8 doi: 10.1006/excr.1996.0052
29	Parsell D A, Lindquist S (1993). The function of heat-shock proteins in stress tolerance: degradation and reactivation of damaged proteins. A Rev Genet , 27: 437–496
30	Pirkkala L, Nykanen P, Sistonen L (2001). Roles of the heat shock transcription factors in regulation of the heat shock response and beyond. FASEB J , 15(7): 1118–1131 doi: 10.1096/fj00-0294rev
31	Qiu Z, Bossier P, Wang X, Bojikova-Fournier S, MacRae T H (2006). Diversity, structure, and expression of the gene for p26, a small heat shock protein from Artemia. Genomics , 88(2): 230–240 doi: 10.1016/j.ygeno.2006.02.008
32	Qiu Z, MacRae T H (2008). ArHsp21, A developmentally regulated small heat-shock protein synthesized in diapausing embryos of Artemia franciscana. Biochem J , 411(3): 605–611 doi: 10.1042/BJ20071472
33	Sun Y, MacRae T H (2005). Small heat shock proteins: molecular structure and chaperone function. Cell Mol Life Sci , 62(21): 2460–2476 doi: 10.1007/s00018-005-5190-4
34	Tanguay R M, Wu Y, Khandjian E W (1993). Tissue-specific expression of heat shock proteins of the mouse in the absence of stress. Dev Genet , 14(2): 112–118 doi: 10.1002/dvg.1020140205
35	Voellmy R (2004). On mechanisms that control heat shock transcription factor activity in metazoan cells. Cell Stress Chaperones , 9(2): 122–133 doi: 10.1379/CSC-14R.1
36	Waters E R, Lee G J, Vierling E (1996). Evolution, structure and function of the small heat shock proteins in plants. J Exp Bot , 47(3): 325– 338 doi: 10.1093/jxb/47.3.325
37	Willsie J K, Clegg J S (2001). Nuclear p26, a small heat shock/α-crystallin protein, and its relationship to stress resistance in Artemia franciscana embryos. J Exp Biol , 204: 2339–2350
38	Willsie J K, Clegg J S (2002). Small heat shock protein p26 associates with nuclear lamins and HSP70 in nuclei and nuclear matrix fractions from stressed cells. J Cell Biochem , 84(3): 601–614 doi: 10.1002/jcb.10040

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