Abstract
Small heat shock proteins (sHSPs) play a crucial role under abiotic stress and are present in all organisms, from eukaryotes to prokaryotes. However, studies on the sHSP gene family in red alga are limited. In this study, we aimed to identify and characterize NysHSP genes from the genome of N. yezoensis, a marine red alga adapted to the stressful intertidal zone. We identified seven NysHSP genes distributed on all three chromosomes. Expression analysis revealed that all NysHSP genes responded to H2O2 and heat stress in the gametophytic thalli, but these genes responded only to heat stress in the sporophytic conchocelis. NysHSP20.3, which has an acidic isoelectric point (pI) and short N-terminal region, was localized as granules in the cytosol. Fluorescence imaging of the NysHSP25.8-GFP and NysHSP28.4-GFP fusion proteins revealed that these proteins were located in the chloroplast. Based on their characteristics and cellular localization, the NysHSPs are divided into two subfamilies. Subfamily I includes four sHSP genes that strongly respond to heat stress and encode a protein localized in the cytosol. The NysHSP gene of subfamily II encodes a polypeptide with a long N-terminal region located in the chloroplast. This study provides insights into the evolution and function of the sHSP gene family of the marine red alga N. yezoensis and how it adapts to the stressful intertidal zone.
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Data Availability
The genome sequencing data of the N, yezoensis strain SG104 used in this study are available and the accession numbers are presented in the Method section.
Change history
15 November 2023
A Correction to this paper has been published: https://doi.org/10.1007/s12192-023-01393-2
References
Abe S, Kurashima A, Yokohama Y, Tanaka J (2001) The cellular ability of desiccation tolerance in Japanese intertidal seaweeds. Bot Mar 44:125–131
Basha E, O’Neill H, Vierling E (2012) Small heat shock proteins and alpha-crystallins: dynamic proteins with flexible functions. Trends Biochem Sci 37:106–117
Blouin NA, Brodie JA, Grossman AC, Xu P, Brawley SH (2011) Porphyra: a marine crop shaped by stress. Trends Plant Sci 16:29–37
Chen ST, He NY, Chen JU, Guo FQ (2017) Identification of core subunits of photosystem II as action sites of HSP21, which is activated by the GUN5-mediated retrograde pathway in Arabidopsis. Plant J 89:1106–1118
Flores-Molina MR, Thomas D, Lovazzano C, Nunez A, Zapata J, Kumar M, Correa JA (2014) Contreras-Porcia, L. Desiccation stress in intertidal seaweeds: effects on morphology, antioxidant responses and photosynthetic performance. Aquat Bot 113:90–99
Gao T, Mo Z, Tang L, Yu X, Du G, Mao Y (2022) Heat shock protein 20 gene superfamilys in red algae: evolutionary and functional diversities. Front Plant Sci 13:817852. https://doi.org/10.3389/fpls.2022.817852
Haslbeck M, Vierling E (2015) A first line of stress defense: small heat shock proteins and their function in protein homeostasis. J Mol Biol 427:1537–1548
Hwang MS, Chung IK, Oh YS (1997) Temperature responses of Porphyra tenera Kjellman and P. yezoensis Ueda (Bangiales, Rhodophyta) from Korea. Algae 12:207–213
Im S, Lee HN, Jung HS, Yang S, Park EJ, Hwang MS, Jeong WJ, Choi DW (2017) Transcriptom-based identification of the desiccation response genes in marine red algae Pyropia tenera (Rhodophyta) and enhancement of abiotic stress tolerance by PtDRG2 in Chlamydomonas. Mar Biotechnol 19:232–245
Jin Y, Yang S, Im S, Jeong WJ, Park EJ, Choi DW (2017) Overexpression of the small heat shock protein, PtsHSP19.3 from marine red algae, Pyropia tenera (Bangiales, Rhodophyta) enhances abiotic stress tolerance in Chlamydomonas. J Plant Biotech 44:287–295
Kobayashi Y, Harada N, Nishimura Y, Saito T, Nakamura M, Fujiwara T, Kuroiwa T, Misumi O (2014) Algae sense exact temperatures: small heat shock proteins are expressed at the survival threshold temperature in Cyanidioschyzon merolae and Chlamydomonas reinhardtii. Genome Biol Evol 7:2731–2740
Lee BH, Won SH, Lee HS, Miyao M, Chung WI, Kim IJ, Jo JK (2000) Expression of the chloroplast-localized small heat shock protein by oxidative stress in rice. Gene 245:283–290
Luo QJ, Zhu ZG, Zhu ZJ, Yang R, Qian FJ, Chen HM, Yan XJ (2014) Different responses to heat shock stress revealed heteromorphic adaptation strategy of Pyropia haitanensis (Bangiales, Rhodophyta). PLoS One 9:e94354. https://doi.org/10.1371/journal.pone.0094354
Ma W, Guan X, Li J, Pan R, Wang L, Liu F, Ma H, Zhu S, Hu J, Ruan YL, Chen X, Zhang T (2019) Mitochondrial small heat shock protein mediates seed germination via thermal sensing. Proc Natl Acad Sci USA 116:4716–4721
McLachlan J (1973) Growth media-marine. In: Stein JR (ed) Handbook of phycological methods. Cambridge University Press, New York, pp 56–60
Neta-Sharir I, Isaacson T, Lurie S, Weiss D (2005) Dual role for tomato heat shock protein 21: protecting photosystem II from oxidative stress and promoting color changes during fruit maturation. Plant Cell 17:1829–1838
Peng LN, Huang LB, Gui TY, Yan XH (2022) Identification and expression profiling of HSP20 gene in Neoporphyra haitanensis. J Appl Phycol. 34:1089–1097. https://doi.org/10.1007/s10811-022-02686-2
Rutgers M, Muranaka LS, Muhlhaus T, Sommer F, Thoms S, Schurig J, Willmund F, Schulz-Raffelt M, Schroda M (2017) Substrates of the chloroplast small heat shock proteins 22E/F point to thermolability as a regulative switch for heat acclimation in Chlamydomonas reinhardtii. Plant Mol Biol 95:579–591
Scharf KD, Siddique M, Vierling E (2001) The expanding family of Arabidopsis thaliana small heat stress proteins and a new family of proteins containing alpha-crystallin domains Acd proteins. Cell Stress Chaperones 6:225–237
Sedaghatmehr M, Mueller-Roeber B, Balazadeh S (2015) The plastid metalloprotease FtH6 and small heat shock protein HSP21 jointly regulate thermomemory in Arabidopsis. Nat Commun 26(7):12439. https://doi.org/10.1038/ncommuns12439
Siddique M, Gernhard S, von Koskull-Doring P, Vierling E, Scharf KD (2008) The plant sHSP superfamily: five new members in Arabidopsis thaliana with unexpected properties. Cell Stress and Chaperones 13:183–197
Sun W, Bernard C, van de Cotte B, van Montagu M, Verbruggen N (2001) At-HSP17.6A, encoding a small heat-shock protein in Arabidopsis, can enhance osmotolerance upon overexpression. Plant J 27:407–415
Takahashi M, Mikami K (2017) Oxidative stress promotes asexual reproduction and apogamy in the red seaweed Pyropia yezoensis. Front Plant Sci 8:62. https://doi.org/10.3389/fpls.2017.00062
Tripathy BC, Oelmüller R (2012) Reactive oxygen species generation and signaling in plants. Plant Signal Behav 7:1621–1633
Uji T, Gondaira Y, Fukuda S, Mizuta H, Saga N (2019) Characterization and expression profiles of small heat shock proteins in the marine red alga Pyropia yezoensis. Cell Stress and Chaperones 24:223–233
Volkov RA, Panchuk II, Schoffl F (2005) Small heat shock proteins are differentially regulated during pollen development and following heat stress in tobacco. Plant Mol Biol 57:487–502
Wang D, Yu X, Xu K, Bi G, Cao M, Zelzion E, Fu C, Sun P, Liu Y, Kong F, Du G, Tang X, Yang R, Wang J, Tang L, Wang L, Zhao Y, Ge Y, Zhuang Y et al (2020) Pyropia yezoensis genome reveals diverse mechanisms of carbon acquisition in the intertidal environment. Nat Commun 12(11):4028. https://doi.org/10.1038/s41467-020-17689-1
Waters ER (2013) The evolution, function, structure, and expression of the plant sHSPs. J Exp Bot 64:391–403
Waters ER, Aevermann BD, Sanders-Reed Z (2008) Comparative analysis of the small heat shock proteins in three angiosperm genomes identifies new subfamilies and reveals diverse evolutionary patterns. Cell Stress Chaperones 13:127–142
Waters ER, Rioflorido I (2007) Evolutionary analysis of the small heat shock proteins in five complete algal genomes. J Mol Evol 65:162–174
Waters ER, Vierling E (1999) Chloroplast small heat shock proteins: evidence for atypical evolution of an organelle-localized protein. Proc Natl Acad Sci USA 96:14394–14399
Waters ER, Vierling E (2020) Plant small heat shock proteins-evolutionary and functional diversity. New Phytol 227:24–37
Yang S, Na Y, Im S, Jo J, Nguyen TD, Kin JS, Jeong WJ, Choi DW (2019) PtsHSP19.6, a small heat-shock protein from the marine red alga Pyropia tenera (Rhodophyta), aggregates into granules and enhances heat tolerance. J Appl Phycol 31:1921–1929
Yu J, Cheng Y, Fen K, Ruan M, Ye Q, Wang R, Li Z, Zhou F, Yao Z, Yang Y, Wang H (2018) Genome-wide identification and expression profiling of tomato HSP20 gene family in response to biotic and abiotic stress. Front Plant Sci 7:1215. https://doi.org/10.3389/fpls.2016.01215
Zhang K, Ezemaduka AN, Wang Z, Hu H, Shi X, Liu C, Lu X, Fu X, Chang Z, Yin CC (2015) A novel mechanism for small heat shock proteins to function as molecular chaperones. Sci Rep 5:8811. https://doi.org/10.1038/srep08811
Zhao P, Wang D, Wang R, Kong N, Zhang C, Yang C, Wu W, Ma H, Chen Q (2018) Genome-wide analysis of the potato HSP20 gene family: identification, genomic organization and expression profiles in response to heat stress. BMG Genom 19:16. https://doi.org/10.1186/s12864-018-4443-1
Zhong L, Zhou W, Wang H, Ding S, Lu Q, Wen X, Peng L, Zhang L, Lu C (2013) Chloroplast small heat shock protein HSP21 interacts with plastid nucleoid protein pTAC5 and is essential for chloroplast development in Arabidopsis under heat stress. Plant Cell 25:2925–2943
Funding
The present study was supported by the National Research Foundation of Korea (NRF) grant (No. 2021R1I1A3047280) funded by the Korean Government; and a grant (No. R2023022) from the National Institute of Fisheries Science, ministry of Oceans and Fisheries, Republic of Korea.
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Supplementary Table 1. The amino acid sequences of the sHSPs included in the phylogenetic analysis. Supplementary Table 2. Comparative threshold (Ct) values of the NysHSP genes in the gametophyte thalli of N. yezoensis under heat, drought stress and H2O2 treatment. Supplementary Table 3. Comparative threshold (Ct) values of the NysHSP genes in the sporophytic conchocelis of N. yezoensis under heat stress, H2O2, and mannitol treatments. (XLSX 25 kb)
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Wi, J., Park, EJ., Hwang, MS. et al. A subfamily of the small heat shock proteins of the marine red alga Neopyropia yezoensis localizes in the chloroplast. Cell Stress and Chaperones 28, 835–846 (2023). https://doi.org/10.1007/s12192-023-01375-4
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DOI: https://doi.org/10.1007/s12192-023-01375-4