J Plant Biotechnol 2017; 44(3): 287-295
Published online September 30, 2017
https://doi.org/10.5010/JPB.2017.44.3.287
© The Korean Society of Plant Biotechnology
Correspondence to : e-mail: dwchoi63@jnu.ac.kr
This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Water temperature is one of the major factors that impacts the growth and life cycle of
Keywords Red algae,
Temperature is a major environmental factor that limit the growth and yield of plant. Under high temperatures or heat stress, plants alter gene expression patterns to adapt to a given environment. Heat shock protein (HSP) genes are key components turned on under heat stress condition and contributing to cellular homeostasis (Scharf et al. 2001; Wang et al. 2004; Schroda and Vallon, 2009; Basha et al. 2012). They are responsible for protein folding, assembly, translocation and degradation in a broad array of normal cellular processes; they also function in the stabilization of proteins and membranes, and can assist in protein refolding under stress conditions. All organism response to high temperature and turning on the HSPs which are a number of conserved protein families such as the HSP100s, HSP90s, HSP70s, HSP60s and sHSP (Wang et al. 2004).
The small heat shock protein (sHSP) is a family of heat shock protein that range in size from approximately 12 ~ 42 kDa. Most sHSPs are in the range of 15 ~ 22 kDa. This protein is characterized by having a conserved C-terminal domain of approximately 90 amino acids referred to as the a-crystallin domain (ACD) (Basha et al. 2012; Scharf et al. 2001; Waters, 2013). Except ACD domain, sHSP are variable in both length and sequence. The sHSPs that function in specific cellular organelles or compartments have N-terminal transit or signal sequences needed to get the sHSP to the proper cellular compartment (Waters, 2013).
The high resolution crystal structures studies show that sHSP are composed of a β-sandwich of two antiparallel sheets and form a hollow ball (van Montfort et al. 2002; Waters, 2013; Zhang et al. 2015). The oligomers are of different sizes. The plant Hsp16.9 from wheat (
The sHSPs are ubiquitously present in all organisms. The number of genes encoding sHSPs in different organisms varies greatly. Investigations of sHSPs from mammals, yeast, plants and bacteria have unveiled their role in thermotolerancesuch that the overexpression of eukaryotic recombinant sHSPscould increase the thermotolerance of
The sHSP are known to act as ATP-independent molecular chaperones work with other chaperones to prevent irreversible aggregations and to re-solubilize proteins that have already aggregated (Waters, 2013; Zhang et al. 2015). They do not require ATP to bind substrate proteins. The sHSP bind to the non-native substrate proteins and release them for refolding with the help of other ATP-dependent chaperones (Waters, 2013). In the absence of such stresses, however, sHSPs can also be produced specifically in reproductive organs at certain developmental stages, including seed maturation and germination, pollen development, and fruit maturation (Neta-sharir et al. 2005; Volkov et al. 2005).
We identified four small heat shock genes (
Transcriptome sequence reads generated from
Sequence editing and amino acid sequence prediction from the selected contigs were conducted using the Sequencherprogram (Gene Code Corp., Ann Arbor, MI, USA). The putative molecular weights and pI values of the deduced polypeptides were predicted using the Compute pI/Mw program (
Gene-specific qRT-PCR was conducted to assay
qRT-PCR was carried out on a Rotor-Gene RG-3000 cycler (Corbett, Sydney, Australia) using the QuantiTect SYBR Green PCR kit (Qiagen), according to the manufacturer’s instructions. The qRT-PCR program consisted of a pre- denaturation step at 95°C for 10 min and 40 cycles of amplification at 95°C for 15 sec, 60°C for 30 sec, and 72°C for 30 sec. All samples were run in duplicate and n-fold differential expression was calculated using the comparative Ct method, 2-ΔΔCt. The
The
The
The transcriptome sequences from
Table 1 Summary of small heat shock proteins (PtsHSPs) isolated from
Gene | contig name | No. amino acid residues | MW (kDa) | pI | e-value | Description |
---|---|---|---|---|---|---|
173 | 19.3 | 5.44 | 6e-96 | HSP22 of | ||
179 | 19.6 | 6.05 | 3e-72 | HSP22 of | ||
200 | 20.2 | 6.42 | 7e-21 | HSP22 of | ||
154 | 16.4 | 4.70 | 1e-19 | HSP of |
Amino acid sequence alignment and phylogenic tree of PtsHSPs isolated from
Except PhsHSP22, there are no known genes showing significant amino acid sequence homology with PtsHSPs in public database. These results indicate that red algae sHSPs are much different from the known sHSPs from green plants including single cell algae and higher land plants (Fig 1B). Or PtsHSP19.3 may have no chaperon activity although ACD domain was found.
A characteristic of many s
Expression pattern of the
To determine the cellular location of the PtsHSP 19.3 protein, the
Subcellular localizations of PtsHSP19.3
A reporter gene encoding green fluorescent protein (GFP) was fused to
Plant have more sHSPs than other eukaryotes (Waters et al. 2008; Yan et al. 2017). Analysis of three angiosperm genome sequence including
Arabidopsis study reported that some cytosolic sHSPs localize as multichaperone complex in cytosol (Siddique et al. 2008). The sHSPs of the cytoplasmic/nuclear subfamilies CI, CII and CIII are shown to be recruited to heat shock complex under heat stress condition (Siddique et al. 2008). Recently Zhang et al (2015) reported that small heat shock protein CeHSP17 from
The complete
Effects of
A. Vector map for the expression of
This work was supported by Korean Institute of Planning and Evaluation for Technology, Agriculture, Forestry and Fisheries (IPET) through Golden Seed Project (Project number, 213008-05-1-SB830), funded by Ministry of Agriculture, Food, and Rural Affairs (MAFRA); the Ministry of Oceans and Fisheries (MOF); the Rural Development Administration (RDA); and the Korea Forest Service (KFS).
J Plant Biotechnol 2017; 44(3): 287-295
Published online September 30, 2017 https://doi.org/10.5010/JPB.2017.44.3.287
Copyright © The Korean Society of Plant Biotechnology.
Yujin Jin, Sungwhan Yang, Sungoh Im, Won-Joong Jeong, EunJeong Park, and Dong-Woog Choi
Department of Biology Education and Kumho Life Science Laboratory, Chonnam National University, Gwangju, 61186, Korea,
Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 34141, Korea,
Seaweed Research Center, National Fisheries Research and Development Institute, Mokpo, 58746, Korea
Correspondence to: e-mail: dwchoi63@jnu.ac.kr
This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Water temperature is one of the major factors that impacts the growth and life cycle of
Keywords: Red algae,
Temperature is a major environmental factor that limit the growth and yield of plant. Under high temperatures or heat stress, plants alter gene expression patterns to adapt to a given environment. Heat shock protein (HSP) genes are key components turned on under heat stress condition and contributing to cellular homeostasis (Scharf et al. 2001; Wang et al. 2004; Schroda and Vallon, 2009; Basha et al. 2012). They are responsible for protein folding, assembly, translocation and degradation in a broad array of normal cellular processes; they also function in the stabilization of proteins and membranes, and can assist in protein refolding under stress conditions. All organism response to high temperature and turning on the HSPs which are a number of conserved protein families such as the HSP100s, HSP90s, HSP70s, HSP60s and sHSP (Wang et al. 2004).
The small heat shock protein (sHSP) is a family of heat shock protein that range in size from approximately 12 ~ 42 kDa. Most sHSPs are in the range of 15 ~ 22 kDa. This protein is characterized by having a conserved C-terminal domain of approximately 90 amino acids referred to as the a-crystallin domain (ACD) (Basha et al. 2012; Scharf et al. 2001; Waters, 2013). Except ACD domain, sHSP are variable in both length and sequence. The sHSPs that function in specific cellular organelles or compartments have N-terminal transit or signal sequences needed to get the sHSP to the proper cellular compartment (Waters, 2013).
The high resolution crystal structures studies show that sHSP are composed of a β-sandwich of two antiparallel sheets and form a hollow ball (van Montfort et al. 2002; Waters, 2013; Zhang et al. 2015). The oligomers are of different sizes. The plant Hsp16.9 from wheat (
The sHSPs are ubiquitously present in all organisms. The number of genes encoding sHSPs in different organisms varies greatly. Investigations of sHSPs from mammals, yeast, plants and bacteria have unveiled their role in thermotolerancesuch that the overexpression of eukaryotic recombinant sHSPscould increase the thermotolerance of
The sHSP are known to act as ATP-independent molecular chaperones work with other chaperones to prevent irreversible aggregations and to re-solubilize proteins that have already aggregated (Waters, 2013; Zhang et al. 2015). They do not require ATP to bind substrate proteins. The sHSP bind to the non-native substrate proteins and release them for refolding with the help of other ATP-dependent chaperones (Waters, 2013). In the absence of such stresses, however, sHSPs can also be produced specifically in reproductive organs at certain developmental stages, including seed maturation and germination, pollen development, and fruit maturation (Neta-sharir et al. 2005; Volkov et al. 2005).
We identified four small heat shock genes (
Transcriptome sequence reads generated from
Sequence editing and amino acid sequence prediction from the selected contigs were conducted using the Sequencherprogram (Gene Code Corp., Ann Arbor, MI, USA). The putative molecular weights and pI values of the deduced polypeptides were predicted using the Compute pI/Mw program (
Gene-specific qRT-PCR was conducted to assay
qRT-PCR was carried out on a Rotor-Gene RG-3000 cycler (Corbett, Sydney, Australia) using the QuantiTect SYBR Green PCR kit (Qiagen), according to the manufacturer’s instructions. The qRT-PCR program consisted of a pre- denaturation step at 95°C for 10 min and 40 cycles of amplification at 95°C for 15 sec, 60°C for 30 sec, and 72°C for 30 sec. All samples were run in duplicate and n-fold differential expression was calculated using the comparative Ct method, 2-ΔΔCt. The
The
The
The transcriptome sequences from
Table 1 . Summary of small heat shock proteins (PtsHSPs) isolated from
Gene | contig name | No. amino acid residues | MW (kDa) | pI | e-value | Description |
---|---|---|---|---|---|---|
173 | 19.3 | 5.44 | 6e-96 | HSP22 of | ||
179 | 19.6 | 6.05 | 3e-72 | HSP22 of | ||
200 | 20.2 | 6.42 | 7e-21 | HSP22 of | ||
154 | 16.4 | 4.70 | 1e-19 | HSP of |
Amino acid sequence alignment and phylogenic tree of PtsHSPs isolated from
Except PhsHSP22, there are no known genes showing significant amino acid sequence homology with PtsHSPs in public database. These results indicate that red algae sHSPs are much different from the known sHSPs from green plants including single cell algae and higher land plants (Fig 1B). Or PtsHSP19.3 may have no chaperon activity although ACD domain was found.
A characteristic of many s
Expression pattern of the
To determine the cellular location of the PtsHSP 19.3 protein, the
Subcellular localizations of PtsHSP19.3
A reporter gene encoding green fluorescent protein (GFP) was fused to
Plant have more sHSPs than other eukaryotes (Waters et al. 2008; Yan et al. 2017). Analysis of three angiosperm genome sequence including
Arabidopsis study reported that some cytosolic sHSPs localize as multichaperone complex in cytosol (Siddique et al. 2008). The sHSPs of the cytoplasmic/nuclear subfamilies CI, CII and CIII are shown to be recruited to heat shock complex under heat stress condition (Siddique et al. 2008). Recently Zhang et al (2015) reported that small heat shock protein CeHSP17 from
The complete
Effects of
A. Vector map for the expression of
This work was supported by Korean Institute of Planning and Evaluation for Technology, Agriculture, Forestry and Fisheries (IPET) through Golden Seed Project (Project number, 213008-05-1-SB830), funded by Ministry of Agriculture, Food, and Rural Affairs (MAFRA); the Ministry of Oceans and Fisheries (MOF); the Rural Development Administration (RDA); and the Korea Forest Service (KFS).
Amino acid sequence alignment and phylogenic tree of PtsHSPs isolated from
Expression pattern of the
Subcellular localizations of PtsHSP19.3
A reporter gene encoding green fluorescent protein (GFP) was fused to
Effects of
A. Vector map for the expression of
Effects of
Table 1 . Summary of small heat shock proteins (PtsHSPs) isolated from
Gene | contig name | No. amino acid residues | MW (kDa) | pI | e-value | Description |
---|---|---|---|---|---|---|
173 | 19.3 | 5.44 | 6e-96 | HSP22 of | ||
179 | 19.6 | 6.05 | 3e-72 | HSP22 of | ||
200 | 20.2 | 6.42 | 7e-21 | HSP22 of | ||
154 | 16.4 | 4.70 | 1e-19 | HSP of |
Journal of
Plant BiotechnologyAmino acid sequence alignment and phylogenic tree of PtsHSPs isolated from
Expression pattern of the
Subcellular localizations of PtsHSP19.3
A reporter gene encoding green fluorescent protein (GFP) was fused to
Effects of
A. Vector map for the expression of
Effects of