RESUMEN
Cysteine-rich transmembrane module (CYSTM) proteins constitute small molecular protein families and have been identified across eukaryotes, including yeast, humans, and several plant species. Plant CYSTMs play vital roles in growth regulation, development, phytohormone signal transduction, pathogen defense, environmental stress response, and even heavy metal binding and detoxification. Canavalia rosea (Sw.) DC is a perennial halophyte with great semi-arid and saline-alkali tolerance. In this study, the CrCYSTM family including 10 members were identified in the C. rosea genome, with the purpose of clarifying the possible roles of CrCYSTMs in C. rosea plants development and stress resistance. The phylogenetic relationships, exon-intron structure, domain structure, chromosomal localization, and putative cis-acting elements in promoter regions were predicted and analyzed. Transcriptome analysis combined with quantitative reverse transcription PCR showed that different CrCYSTM members exhibited varied expression patterns in different tissues and under different abiotic stress challenges. In addition, several CrCYSTMs were cloned and functionally characterized for their roles in abiotic stress tolerance with yeast expression system. Overall, these findings provide a foundation for functionally characterizing plant CYSTMs to unravel their possible roles in the adaptation of C. rosea to tropical coral reefs. Our results also lay the foundation for further research on the roles of plant CYSTM genes in abiotic stress signaling, especially for heavy metal detoxification.
Asunto(s)
Canavalia , Cisteína , Humanos , Cisteína/metabolismo , Canavalia/genética , Canavalia/metabolismo , Saccharomyces cerevisiae/metabolismo , Filogenia , Estrés Fisiológico/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas , Familia de MultigenesRESUMEN
Objectives: Angiotensin II (Ang II) plays a key role in the regulation of myocardial hypertrophy via downstream cysteine-rich transmembrane bone morphogenetic protein regulator 1 (Crim1). However, it is still unclear whether Crim1 is involved in ionic channel remodeling. The study aimed to explore the effects of Crim1 on transient outward potassium current (Ito) and Kv4.2 (the main subunit of Ito channel) expression in hypertrophic ventricular cardiomyocytes. Materials and Methods: The ventricular cardiomyocytes were isolated from the neonatal rats. Hypertrophy was induced by Ang II. Crim1 expression was modulated by using adenovirus transfection. The expression of myosin heavy chain beta (ß-MHC), Crim1, and Kv4.2 was determined by RT-qPCR and western blot. The cellular surface area was assessed using Image J software. Ito was recorded by the whole-cell patch clamp technique. Results: Ang II-induced hypertrophy in cardiomyocytes was identified by their larger cellular surface area and higher mRNA expression of ß-MHC. Ang II significantly decreased the expression of Crim1 and Kv4.2 and reduced Ito current density. However, Crim1 overexpression abolished the Ang II-induced hypertrophy and preserved the expression of Kv4.2 and Ito current density. Conclusion: Crim1 overexpression inhibits Ang II-induced hypertrophy and preserves Ito current density via up-regulating Kv4.2 in ventricular cardiomyocytes from neonatal rats. Crim1 could have a role in the development of ventricular arrhythmia in hypertrophic hearts.
RESUMEN
Left ventricular hypertrophy is a leading cause of heart failure and sudden death. Cysteine-rich transmembrane bone morphogenetic protein regulator 1 (Crim1) is expressed at a high level in the heart and has a regulatory role in heart development. The present study aimed to test the hypothesis that Crim1 can have an inhibitory function on ventricular hypertrophy. Rat primary ventricular myocytes were stretched to induce myocyte hypertrophy, and treated with telmisartan or infected with Crim1-expressing recombinant adenovirus (Ad-Crim1). Rat ventricular hypertrophy was induced by abdominal aortic coarctation (AAC), and treated either with telmisartan or myocardial injection of Ad-Crim1 or empty adenovirus vector. The results showed that the expression of Crim1 decreased in the hypertrophic ventricle. The inhibition of angiotensin receptor type 1 (AT1R) by telmisartan in vitro and in vivo significantly increased the expression of Crim1 in the left ventricle. The overexpression of Crim1 by infection with Ad-Crim1 significantly inhibited stretch-induced ventricular myocyte hypertrophy in vitro. The overexpression of Crim1 by gavage with AT1R inhibitor telmisartan or myocardial injection of Ad-Crim1 markedly suppressed AAC-induced left ventricular hypertrophy in vivo. These results suggest that Crim1 has a suppressive function on ventricular hypertrophy and provides a novel therapeutic target for the treatment of cardiac hypertrophy.
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Objective To study the expression and the clinical significance of cysteine rich transmembrane BMP regulator 1 (CRIM1) in hepatocellular carcinoma (HCC) and discuss the association between CRIM1 and epithelial-mesenchymal transition (EMT).Methods The cases were came from the Subei People's Hospital Affilated Hospital of Yangzhou University from January 2013 to December 2017.CRIM1 and EMT related proteins (E-cadherin,Vimentin) in parts of HCC tissues and their paired peritumoural tissues were tested by Western blotting.The gray value was test by t test.The observation indicators:(1) expression of CRIM1 protein and EMT-related protein (E-cadherin,Vimentin) in liver cancer tissues and paracancerous tissues.(2) The relationship between CRIM1 protein expression and clinicopathological factors in patients with liver cancer.The expression ofCRIM1 in HCC tissues and adjacent tissues was detected by immunohistochemistry(IHC),which was divided into high expression group and low expression group according to the histochemical score,and the relation between the expression of CRIM1 and the clinicopathological factors of the patients was analyzed by chi-square test and Spearman correlation analysis.Finally,the relation between CRIM1 and overall survival of HCC patients was analyzed by Kaplan Meier Plotter database.Results The expression of CRIM1 in tumor and matched paratumor specimens were 0.15 ± 0.03,0.8 ± 0.04,and E-cadherinin tumor and matched paratumor specimenswere 0.20 ±0.05,0.56 ± 0.06,their expression in paracancerous tissues was higher than HCC tissues (t =14.21,4.69,P ＜ 0.05),while the expression of Vimentin in tumor and matched paratumor specimens were 0.74 ± 0.08,0.45 ± 0.06,the expression in tumor tissues were significantly higher than adjacent tissues (t =2.87,P ＜ 0.05).The expression of CRIM1 in HCC tissues was further verified by immunohistochemistry,which shows that CRIM1 was overexpressed in paracancerous tissues.In 114 patients,46 cases of CRIM1 protein were highly expressed in liver cancer tissues,and 68 cases of CRIM1 protein were low expressed.The expression of CRIM1 obviously related with the level of αt-fetoprotein (AFP),tumor size and symptom(r =-0.43,-0.34,-0.24,x2 =9.381,5.248,8.117,P ＜ 0.05).However,other clinicopathological features were not correlated with CRIM 1 expression,including age,tumour differentiation,tumor number.Finally,the overall survival was different between CRIM1 high expression and low expression according to the Kaplan Meier Plotter database.Conclusions The expression of CRIM1 is negatively correlated with the EMT process in HCC.CRIM1 might be a potential molecular marker for prognosis.
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The developing lens is a powerful system for investigating the molecular basis of inductive tissue interactions and for studying cataract, the leading cause of blindness. The formation of tightly controlled cell-cell adhesions and cell-matrix junctions between lens epithelial (LE) cells, between lens fiber (LF) cells, and between these two cell populations enables the vertebrate lens to adopt a highly ordered structure and acquire optical transparency. Adhesion molecules are thought to maintain this ordered structure, but little is known about their identity or interactions. Cysteine-rich motor neuron 1 (Crim1), a type I transmembrane protein, is strongly expressed in the developing lens and its mutation causes ocular disease in both mice and humans. How Crim1 regulates lens morphogenesis is not understood. We identified a novel ENU-induced hypomorphic allele of Crim1, Crim1(glcr11), which in the homozygous state causes cataract and microphthalmia. Using this and two other mutant alleles, Crim1(null) and Crim1(cko), we show that the lens defects in Crim1 mouse mutants originate from defective LE cell polarity, proliferation and cell adhesion. Crim1 adhesive function is likely to be required for interactions both between LE cells and between LE and LF cells. We show that Crim1 acts in LE cells, where it colocalizes with and regulates the levels of active ß1 integrin and of phosphorylated FAK and ERK. The RGD and transmembrane motifs of Crim1 are required for regulating FAK phosphorylation. These results identify an important function for Crim1 in the regulation of integrin- and FAK-mediated LE cell adhesion during lens development.