Type II Ice-Binding Proteins Isolated from an Arctic Microalga Are Similar to Adhesin-Like Proteins and Increase Freezing Tolerance in Transgenic Plants

2019 ◽  
Vol 60 (12) ◽  
pp. 2744-2757 ◽  
Author(s):  
Sung Mi Cho ◽  
Sanghee Kim ◽  
Hojin Cho ◽  
Hyoungseok Lee ◽  
Jun Hyuck Lee ◽  
...  
Keyword(s):  
Low Temperature ◽  
Transgenic Plants ◽  
Freezing Tolerance ◽  
Binding Proteins ◽  
The Arctic ◽  
Ice Recrystallization Inhibition ◽  
Ice Binding ◽  
Antarctic Snow ◽  
Vitro Analysis

Abstract Microalgal ice-binding proteins (IBPs) in the polar region are poorly understood at the genome-wide level, although they are important for cold adaptation. Through the transcriptome study with the Arctic green alga Chloromonas sp. KNF0032, we identified six Chloromonas IBP genes (CmIBPs), homologous with the previously reported IBPs from Antarctic snow alga CCMP681 and Antarctic Chloromonas sp. They were organized with multiple exon/intron structures and low-temperature-responsive cis-elements in their promoters and abundantly expressed at low temperature. The biological functions of three representative CmIBPs (CmIBP1, CmIBP2 and CmIBP3) were tested using in vitro analysis and transgenic plant system. CmIBP1 had the most effective ice recrystallization inhibition (IRI) activities in both in vitro and transgenic plants, and CmIBP2 and CmIBP3 had followed. All transgenic plants grown under nonacclimated condition were freezing tolerant, and especially 35S::CmIBP1 plants were most effective. After cold acclimation, only 35S::CmIBP2 plants showed slightly increased freezing tolerance. Structurally, the CmIBPs were predicted to have β-solenoid forms with parallel β-sheets and repeated TXT motifs. The repeated TXT structure of CmIBPs appears similar to the AidA domain-containing adhesin-like proteins from methanogens. We have shown that the AidA domain has IRI activity as CmIBPs and phylogenetic analysis also supported that the AidA domains are monophyletic with ice-binding domain of CmIBPs, and these results suggest that CmIBPs are a type of modified adhesins.

Biochemical identification of the OsMKK6–OsMPK3 signalling pathway for chilling stress tolerance in rice1

2012 ◽  
Vol 443 (1) ◽  
pp. 95-102 ◽  
Author(s):  
Guosheng Xie ◽  
Hideki Kato ◽  
Ryozo Imai
Keyword(s):  
Protein Kinase ◽  
Low Temperature ◽  
Transgenic Plants ◽  
Stress Tolerance ◽  
Chilling Stress ◽  
Active Form ◽  
Signalling Pathway ◽  
Kinase Assay ◽  
Mapk Kinase

MAPK (mitogen-activated protein kinase) pathways have been implicated in stress signalling in plants. In the present study, we performed yeast two-hybrid screening to identify partner MAPKs for OsMKK (Oryza sativa MAPK kinase) 6, a rice MAPK kinase, and revealed specific interactions of OsMKK6 with OsMPK3 and OsMPK6. OsMPK3 and OsMPK6 each co-immunoprecipitated OsMKK6, and both were directly phosphorylated by OsMKK6 in vitro. An MBP (myelin basic protein) kinase assay of the immunoprecipitation complex indicated that OsMPK3 and OsMPK6 were activated in response to a moderately low temperature (12°C), but not a severely low temperature (4°C) in rice seedlings. A constitutively active form of OsMKK6, OsMKK6DD, showed elevated phosphorylation activity against OsMPK3 and OsMPK6 in vitro. OsMPK3, but not OsMPK6, was constitutively activated in transgenic plants overexpressing OsMKK6DD, indicating that OsMPK3 is an in vivo target of OsMKK6. Enhanced chilling tolerance was observed in the transgenic plants overexpressing OsMKK6DD. Taken together, our data suggest that OsMKK6 and OsMPK3 constitute a moderately low-temperature signalling pathway and regulate cold stress tolerance in rice.

scholarly journals Protection of Alcohol Dehydrogenase against Freeze–Thaw Stress by Ice-Binding Proteins Is Proportional to Their Ice Recrystallization Inhibition Property

Marine Drugs ◽  
2020 ◽  
Vol 18 (12) ◽  
pp. 638
Author(s):  
Young Hoon Lee ◽  
Kitae Kim ◽  
Jun Hyuck Lee ◽  
Hak Jun Kim
Keyword(s):  
Alcohol Dehydrogenase ◽  
Binding Proteins ◽  
Residual Activity ◽  
Fluorescence Emission ◽  
Fluorescence Emission Spectra ◽  
Ice Recrystallization Inhibition ◽  
Ice Recrystallization ◽  
Freeze Thaw ◽  
Ice Binding ◽  
Recrystallization Inhibition

Ice-binding proteins (IBPs) have ice recrystallization inhibition (IRI) activity. IRI property has been extensively utilized for the cryopreservation of different types of cells and tissues. Recent reports demonstrated that IRI can also play a significant role in protecting proteins from freezing damage during freeze–thaw cycles. In this study, we hypothesized that the protective capability of IBPs on proteins against freeze–thaw damage is proportional to their IRI activity. Hence we used two IBPs: one with higher IRI activity (LeIBP) and the other with lower activity (FfIBP). Yeast alcohol dehydrogenase (ADH) was used as a freeze-labile model protein. IBPs and ADH were mixed, frozen at −20 °C, and thawed repeatedly. The structure of ADH was assessed using fluorescence emission spectra probed by 1-anilinonaphthalene-8-sulfonate over the repeated freeze–thaw cycles. The activity was monitored at 340 nm spectrophotometrically. Fluorescence data and activity clearly indicated that ADH without IBP was freeze-labile. However, ADH maintained about 70% residual activity after five repeated cycles at a minimal concentration of 0.1 mg mL-1 of high IRI-active LeIBP, but only 50% activity at 4 mg mL−1 of low active FfIBP. These results showed that the protection of proteins from freeze–thaw stress by IBPs is proportional to their IRI activity.

Gene expression during cold and heat shock in wheat

1991 ◽  
Vol 69 (5-6) ◽  
pp. 383-391 ◽  
Author(s):  
Jean Danyluk ◽  
Eric Rassart ◽  
Fathey Sarhan
Keyword(s):  
Heat Shock ◽  
Low Temperature ◽  
Winter Wheat ◽  
Freezing Tolerance ◽  
Spring Wheat ◽  
Triticum Aestivum L ◽  
Messenger Rnas ◽  
Free System ◽  
Heat And Cold Stress

Translatable messenger RNAs expression was compared in cold- and heat-stressed winter wheat (Triticum aestivum L. 'Fredrick' and 'Norstar') and spring wheat (T. aestivum L. 'Glenlea'). Polyadenylated RNA isolated from the crown and leaf tissues was translated in a wheat germ cell free system and the acidic and basic in vitro products were resolved by two-dimensional SDS–PAGE and autoradiography. The results showed that low temperature stress rapidly induced two groups of mRNAs. The first group was transient in nature and consists of 18 mRNAs that reached their highest levels of induction after 24 h of low temperature exposure and then decreased to undetectable levels. The second group consists of 53 mRNAs that were also induced or increased rapidly, but maintained their levels of expression during the 4 weeks required to induce freezing tolerance. Among those, at least 34 were expressed at higher levels in the freezing tolerant winter wheat compared with the less tolerant spring wheat. This suggests a possible relation between the expression of these mRNAs and the capacity of each genotype to develop freezing tolerance. In the case of heat shock, 50 mRNAs were induced or increased after 3 h at 40 °C. Among these, the expression of only six mRNAs was altered in a similar manner in the three genotypes by both treatments. The remaining mRNAs code for typical heat shock proteins which are different from those induced by low temperature. None of these mRNAs has been associated with the development of freezing tolerance. These results suggest that heat and cold stress are controlled by different genetic systems.Key words: wheat, mRNAs, proteins, low temperature, heat stress.

Expression of CP:CBF3-35S:ICE1 Enhances Low Temperature Tolerance in Transgenic Arabidopsis

2013 ◽  
Vol 726-731 ◽  
pp. 118-121
Author(s):  
Rui Mei Li ◽  
Du Juan Xi ◽  
Yi Meng Ji ◽  
Rui Jun Duan ◽  
Jiao Liu ◽  
...  
Keyword(s):  
Low Temperature ◽  
Transgenic Plants ◽  
Temperature Stress ◽  
Target Genes ◽  
Temperature Tolerance ◽  
Low Temperature Stress ◽  
Stress Tests ◽  
Low Temperature Tolerance ◽  
Transgenic Lines

We have constructed a vector pCAMBIA1300-CP:CBF3-35S:ICE1 and transformed into Arabidopsis. Results of PCR proved that the target genes had integrated into Arabidopsis genome. Transgenic Arabidopsis showed a bit slow growth, earlier flowering, but normal at other phenotype under 22°C with 8 h daily lights. In vitro low temperature stress tests showed that the transgenic lines were survival while the wild type was nearly dead. The transgenic plants also showed an increased proline content, SOD and POD activities under low temperature stress. The phenotype and physical evidence indicated that expression of CP:CBF3-35S:ICE1 under low temperature enhances the cold tolerance in transgenic plants.

scholarly journals Ice Binding Proteins: Diverse Biological Roles and Applications in Different Types of Industry

Biomolecules ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 274 ◽  
Author(s):  
Aneta Białkowska ◽  
Edyta Majewska ◽  
Aleksandra Olczak ◽  
Aleksandra Twarda-Clapa
Keyword(s):  
Binding Proteins ◽  
Thermal Hysteresis ◽  
Industrial Applications ◽  
Food Storage ◽  
Ice Recrystallization Inhibition ◽  
Ice Recrystallization ◽  
Current State ◽  
Ice Binding ◽  
Recrystallization Inhibition ◽  
Living Organisms

More than 80% of Earth’s surface is exposed periodically or continuously to temperatures below 5 °C. Organisms that can live in these areas are called psychrophilic or psychrotolerant. They have evolved many adaptations that allow them to survive low temperatures. One of the most interesting modifications is production of specific substances that prevent living organisms from freezing. Psychrophiles can synthesize special peptides and proteins that modulate the growth of ice crystals and are generally called ice binding proteins (IBPs). Among them, antifreeze proteins (AFPs) inhibit the formation of large ice grains inside the cells that may damage cellular organelles or cause cell death. AFPs, with their unique properties of thermal hysteresis (TH) and ice recrystallization inhibition (IRI), have become one of the promising tools in industrial applications like cryobiology, food storage, and others. Attention of the industry was also caught by another group of IBPs exhibiting a different activity—ice-nucleating proteins (INPs). This review summarizes the current state of art and possible utilizations of the large group of IBPs.

scholarly journals Induction of methyl-CpG-binding domain proteins in coronavirus infection.

2020 ◽  
Author(s):  
Shahan Mamoor
Keyword(s):  
Microarray Data ◽  
Binding Proteins ◽  
Significant Differential Expression ◽  
Coronavirus Infection ◽  
Human Coronaviruses ◽  
Vitro Analysis ◽  
Transcriptional Induction ◽  
In Vitro Analysis

Coronavirus SARS-CoV-2 (“COVID-19”) has infected close to 20,000,000 people worldwide (1, 2). We mined published and public microarray data (3-8) to determine in an unbiased fashion genes most transcriptionally perturbed in the host following coronavirus infection with a series of human coronaviruses, including SARS-CoV-1, MERS-CoV and HCoV-229E. We observed significant transcriptional induction of the methyl-CpG-binding proteins MBD5 and MeCP2 following infection with two genetically distinct MERS-CoV isolates in human cells in vitro. Analysis of independent microarray data revealed less significant differential expression of MBD5 and MeCP2 in the lungs of mice following infection with SARS-CoV-1 in vivo. Epigenetic mechanisms involving methyl-CpG-binding proteins may be important to establishment of coronavirus infection in the human host.

scholarly journals Ice-binding proteins confer freezing tolerance in transgenicArabidopsis thaliana

2016 ◽  
Vol 15 (1) ◽  
pp. 68-81 ◽  
Author(s):  
Melissa Bredow ◽  
Barbara Vanderbeld ◽  
Virginia K. Walker
Keyword(s):  
Freezing Tolerance ◽  
Binding Proteins ◽  
Ice Binding

The native structure of the eukaryotic ribosome

1983 ◽  
Vol 41 ◽  
pp. 432-433
Author(s):  
R.A. Milligan ◽  
P.N.T. Unwin
Keyword(s):  
Ribosomal Proteins ◽  
Crystal Form ◽  
Native Structure ◽  
X Ray Diffraction ◽  
Eukaryotic Ribosome ◽  
Vitro Analysis ◽  
In Vitro Analysis ◽  
Resolution Structure ◽  
Stable Unit

A detailed understanding of the mechanism of protein synthesis will ultimately depend on knowledge of the native structure of the ribosome. Towards this end we have investigated the low resolution structure of the eukaryotic ribosome embedded in frozen buffer, making use of a system in which the ribosomes crystallize naturally.The ribosomes in the cells of early chicken embryos form crystalline arrays when the embryos are cooled at 4°C. We have developed methods to isolate the stable unit of these arrays, the ribosome tetramer, and have determined conditions for the growth of two-dimensional crystals in vitro, Analysis of the proteins in the crystals by 2-D gel electrophoresis demonstrates the presence of all ribosomal proteins normally found in polysomes. There are in addition, four proteins which may facilitate crystallization. The crystals are built from two oppositely facing P4 layers and the predominant crystal form, accounting for >80% of the crystals, has the tetragonal space group P4212, X-ray diffraction of crystal pellets demonstrates that crystalline order extends to ~ 60Å.

1163: Radial Dilation of Ureteral Balloons: A Comparative in-Vitro Analysis

2005 ◽  
Vol 173 (4S) ◽  
pp. 315-316
Author(s):  
Kari Hendlin ◽  
Brynn Lund ◽  
Manoj Monga
Keyword(s):  
Vitro Analysis ◽  
In Vitro Analysis
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