2.Characterization the response of Chlamydomonas reinhardtii serine/threonine protein kinase mutant to blue light.
Wangning LI ; Mengjing LIANG ; Ze YANG ; Yanan LI ; Chunhui ZHANG ; Chunli JI ; Runzhi LI ; Song QIN ; Jinai XUE ; Hongli CUI
Chinese Journal of Biotechnology 2023;39(11):4563-4579
In order to investigate the molecular mechanism of silk/threonine protein kinase (STK)-mediated blue light response in the algal Chlamydomonas reinhardtii, phenotype identification and transcriptome analysis were conducted for C. reinhardtii STK mutant strain crstk11 (with an AphvIII box reverse insertion in stk11 gene coding region) under blue light stress. Phenotypic examination showed that under normal light (white light), there was a slight difference in growth and pigment contents between the wild-type strain CC5325 and the mutant strain crstk11. Blue light inhibited the growth and chlorophyll synthesis in crstk11 cells, but significantly promoted the accumulation of carotenoids in crstk11. Transcriptome analysis showed that 860 differential expression genes (DEG) (559 up-regulated and 301 down-regulated) were detected in mutant (STK4) vs. wild type (WT4) upon treatment under high intensity blue light for 4 days. After being treated under high intensity blue light for 8 days, a total of 1 088 DEGs (468 upregulated and 620 downregulated) were obtained in STK8 vs. WT8. KEGG enrichment analysis revealed that compared to CC5325, the crstk11 blue light responsive genes were mainly involved in catalytic activity of intracellular photosynthesis, carbon metabolism, and pigment synthesis. Among them, upregulated genes included psaA, psaB, and psaC, psbA, psbB, psbC, psbD, psbH, and L, petA, petB, and petD, as well as genes encoding ATP synthase α, β and c subunits. Downregulated genes included petF and petJ. The present study uncovered that the protein kinase CrSTK11 of C. reinhardtii may participate in the blue light response of algal cells by mediating photosynthesis as well as pigment and carbon metabolism, providing new knowledge for in-depth analysis of the mechanism of light stress resistance in the algae.
Chlamydomonas reinhardtii/genetics*
;
Photosynthesis/genetics*
;
Plants/metabolism*
;
Protein Kinases
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Threonine/metabolism*
;
Carbon/metabolism*
;
Serine/metabolism*
3.Production of L-2-aminobutyric acid from L-threonine using a trienzyme cascade.
Yan FU ; Junxuan ZHANG ; Xuerong FU ; Yuchen XIE ; Hongyu REN ; Jia LIU ; Xiulai CHEN ; Liming LIU
Chinese Journal of Biotechnology 2020;36(4):782-791
L-2-aminobutyric acid (L-ABA) is an important chemical raw material and chiral pharmaceutical intermediate. The aim of this study was to develop an efficient method for L-ABA production from L-threonine using a trienzyme cascade route with Threonine deaminase (TD) from Escherichia. coli, Leucine dehydrogenase (LDH) from Bacillus thuringiensis and Formate dehydrogenase (FDH) from Candida boidinii. In order to simplify the production process, the activity ratio of TD, LDH and FDH was 1:1:0.2 after combining different activity ratios in the system in vitro. The above ratio was achieved in the recombinant strain E. coli 3FT+L. Moreover, the transformation conditions were optimized. Finally, we achieved L-ABA production of 68.5 g/L with a conversion rate of 99.0% for 12 h in a 30-L bioreactor by whole-cell catalyst. The environmentally safe and efficient process route represents a promising strategy for large-scale L-ABA production in the future.
Aminobutyrates
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chemical synthesis
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Bacillus thuringiensis
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enzymology
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Candida
;
enzymology
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Escherichia coli
;
enzymology
;
Formate Dehydrogenases
;
metabolism
;
Leucine Dehydrogenase
;
metabolism
;
Threonine
;
metabolism
;
Threonine Dehydratase
;
metabolism
5.Recent advances of studies on role of mTOR signaling in aging of hematopoietic and other organ systems-review.
Chun-Lan HUA ; Tao CHENG ; Wei-Ping YUAN
Journal of Experimental Hematology 2013;21(5):1313-1317
Mammalian target of rapamycin (mTOR) is a serine/threonine protein kinase, which plays an essential role in cell growth, proliferation and survival. mTOR regulates the transcription of mRNA, synthesis of ribosome and gene expression for metabolism. By forming mTOR complex, it regulates cellular activities by phosphorylating its downstream proteins, such as S6 protein kinase and 4E-BP1. In recent years, the role of mTORC1 in regulating aging is gradually recognized. Studies of physiological function and the regulatory mechanisms of mTOR signaling can not only help to better understand the aging mechanism for cells or organs, but also provide insights as to finding potential new drug targets for aging related diseases. This review focuses on recent advances of mTOR and aging related diseases in hematopoietic and other organ systems.
Aging
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Hematopoietic System
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metabolism
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Humans
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Signal Transduction
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TOR Serine-Threonine Kinases
;
metabolism
7.ubtor Mutation Causes Motor Hyperactivity by Activating mTOR Signaling in Zebrafish.
Tiantian WANG ; Mingshan ZHOU ; Quan ZHANG ; Cuizhen ZHANG ; Gang PENG
Neuroscience Bulletin 2021;37(12):1658-1670
Mechanistic target of rapamycin (mTOR) signaling governs important physiological and pathological processes key to cellular life. Loss of mTOR negative regulators and subsequent over-activation of mTOR signaling are major causes underlying epileptic encephalopathy. Our previous studies showed that UBTOR/KIAA1024/MINAR1 acts as a negative regulator of mTOR signaling, but whether UBTOR plays a role in neurological diseases remains largely unknown. We therefore examined a zebrafish model and found that ubtor disruption caused increased spontaneous embryonic movement and neuronal activity in spinal interneurons, as well as the expected hyperactivation of mTOR signaling in early zebrafish embryos. In addition, mutant ubtor larvae showed increased sensitivity to the convulsant pentylenetetrazol, and both the motor activity and the neuronal activity were up-regulated. These phenotypic abnormalities in zebrafish embryos and larvae were rescued by treatment with the mTORC1 inhibitor rapamycin. Taken together, our findings show that ubtor regulates motor hyperactivity and epilepsy-like behaviors by elevating neuronal activity and activating mTOR signaling.
Animals
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Hyperkinesis/genetics*
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Mutation/genetics*
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Signal Transduction
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TOR Serine-Threonine Kinases/metabolism*
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Zebrafish/metabolism*
9.Expression of integrin-linked kinase in oral squamous cell cancinoma and its significance.
Chinese Journal of Stomatology 2010;45(10):623-626
OBJECTIVETo investigate the expression patterns of integrin-linked kinase (ILK) in oral squamous cell carcinoma (OSCC).
METHODSThe expression of ILK in 170 OSCC cases was examined by immunohistochemistry and periodic acid-schiff histochemistry.
RESULTSILK immunoreactivity occurred in cancer cells and(or) stroma encircling the malignant tissue. Along with the deteriorated histopathologic grade of oral cancer, there was stepwise aberration exhibiting from ILK-negative, to ILK-positive in epithelia, to ILK-positive in both epithelia and stroma, and to ILK-positive in stroma. Through non-parameter test, in both lymphatic invasion [ILK-positive in stroma in well-differentiated, moderately-differentiated and poorly-differentiated is 2% (1/50), 18% (6/33) and 64% (18/28), respectively] and non-lymphatic invasion group [ILK-positive in stroma in well-differentiated, moderately-differentiated and poorly-differentiated is 16% (4/25), 9/18 and 10/16, respectively), the expression patterns of ILK exhibited significant correlations with histopathology (F = 17.742, P < 0.001, F = 4.394, P = 0.017). ILK expression between lymphatic metastasis and non-lymphatic metastasis group was significantly different (χ(2) = 14.418, P = 0.002).
CONCLUSIONSILK inclines to express in stroma during the OSCC progress.
Carcinoma, Squamous Cell ; metabolism ; Cell Differentiation ; Epithelial Cells ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Mouth Neoplasms ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism
10.The changes of the expression of 2-type small conductance-Ca- activating-K (SK2) channel protein in hypertensive rat myocardium.
Wen-Fei ZHANG ; Chang-Zhen YANG ; Peng-Cheng HU ; Hao CHEN ; Yue XI ; Hong-Kun FAN ; Qian ZHANG ; Chun YANG
Chinese Journal of Applied Physiology 2019;35(4):381-384
OBJECTIVE:
To investigate the expression of 2-type small conductance-Ca-activating-K (SK2) channel protein in hypertensive rat myocardial cells.
METHODS:
Twelve healthy adult male SD rats were randomly divided into control group (n=5) and experimental group (n=7). The rats of experimental group were injected intraperitoneally with N'-nitro-L-arginine (L-NNA 15 mg/(kg·d))while the rats of control group were injected intraperitoneally with isometrical normal saline(15 ml/(kg·d )). The body weight, blood pressure and electrocardiogram of the rats were measured every week. After 4 weeks, the rats were sacrificed to obtain hearts, and the expression of SK2 channel protein in myocardium was detected by Western blot.
RESULTS:
After 4 weeks of administration, compared with the control group, the blood pressure in the experimental group was significantly elevated (P<0.05), QRS duration and R-R interval were prolonged, and the expressions of SK2 channel in the atrial and ventricular tissue of the experimental group were significantly higher than those in the control group (1.12±0.18,1.64±0.26, P < 0.05).
CONCLUSION
The expressions of atrial and ventricular SK2 pathway are increased in hypertensive model rats. It may be one of the mechanism leading to arrhythmias in hypertensive model rats and can provide new ideas and strategies for the treatment and prognosis of hypertensive diseases.
Animals
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Hypertension
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metabolism
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Male
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Myocardium
;
metabolism
;
Protein-Serine-Threonine Kinases
;
metabolism
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Random Allocation
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Rats
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Rats, Sprague-Dawley