• Title/Summary/Keyword: Actin mRNA

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Regulation of Actin Gene Expression During the Differentiation of Naegleria gruberi

  • Kim, Misook;Lee, Joo-Hun
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.42-48
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    • 2001
  • The regulation of actin gene expression during the differentiation of Naegleria gruberi was examined. Actin mRNA concentration was maximal in amoebae and decreased rapidly after the initiation of differentiation. At 20 min after initiation, the concentration of actin mRNA decreased to 55% of the maximal value. The actin mRNA concentration decreased to the minimum at 80 min (15% of the maximum), and then began to increase slightly at the end of differentiation. This decrease of actin mRNA concentration was regulated by the repression of actin gene transcription based on nuclear run-on transcription experiments. The rates of transcription of actin gene in nuclei prepared at 40 and 80 min after the initiation of differentiation were 50 and 28% of that of nuclei prepared at the beginning of differentiation, respectively. The addition of cycloheximide at the initiation of differentiation inhibited both the rapid decrease in the concentration of actin mRNA and the repression of actin gene transcription. These results suggest that the rapid decrease in the concentration of actin mRNA during the differentiation of N. gruberi is accomplished by the repression of actin gene transcription and this transcriptional regulation requires continuous protein synthesis during the differentiation.

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Effect of 'Sexiang Shuhuo Jing' for CPK, LDH Activities and Skeletal Muscle ${\alpha}-actin$ mRNA Expression after Skeletal Muscle in Rats (골격근 손상에 대한 '사향서활정'(麝香舒活精)치료의 혈청 CK, LDH활성도 및 골격근 ${\alpha}-actin$ mRNA 발현 변화의 관찰)

  • Kim, Jin-Hang;Song, Je-Ho
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.4
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    • pp.992-996
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    • 2006
  • The study examined clinical effect of the 'Sexiang Shuhuo Jing' on serum CK and LDH activities and skeletal muscle ${\alpha}-actin$ mRNA expression concentration 140days after skeletal muscle injury in rats. The clinical research consisted of observing and measuring the serum CK, LDH activities and skeletal muscle ${\alpha}-actin$ mRNA expression, at the time of injury and during recovery. All experimental data were analyzed by repeated measurement with ANOVA on of SPSS(11.5v), accepting level for all significances was above ${\alpha}\;=.05.$ The results were as follows: That skeletal muscle injury in rats there existed a substantial increase serum CK, LDH activities and expression of skeletal muscle ${\alpha}-actin$ mRNA And Sexiang Shuhuo Jing treatment group's serum CK, LDH activities lower and faster recovery than control group. The 1 st day after skeletal muscle injury, Sexiang Shuhuo Jing treatment group's skeletal muscle ${\alpha}-actin$ mRNA expression was much more higher than control group, after 2 day's faster recovery normal level than control group. There existed a substantial increase again serum CK, LDH activities and skeletal muscle ${\alpha}-actin$ mRNA expression 3rd days after injury in control group. But in Sexiang Shuhuo Jing treatment group's can't be found that.

MS2 Labeling of Endogenous Beta-Actin mRNA Does Not Result in Stabilization of Degradation Intermediates

  • Kim, Songhee H.;Vieira, Melissa;Kim, Hye-Jin;Kesawat, Mahipal Singh;Park, Hye Yoon
    • Molecules and Cells
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    • v.42 no.4
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    • pp.356-362
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    • 2019
  • The binding of MS2 bacteriophage coat protein (MCP) to MS2 binding site (MBS) RNA stem-loop sequences has been widely used to label mRNA for live-cell imaging at single-molecule resolution. However, concerns have been raised recently from studies with budding yeast showing aberrant mRNA metabolism following the MS2-GFP labeling. To investigate the degradation pattern of MS2-GFP-labeled mRNA in mammalian cells and tissues, we used Northern blot analysis of ${\beta}$-actin mRNA extracted from the Actb-MBS knock-in and $MBS{\times}MCP$ hybrid mouse models. In the immortalized mouse embryonic cell lines and various organ tissues derived from the mouse models, we found no noticeable accumulation of decay products of ${\beta}$-actin mRNA compared with the wild-type mice. Our results suggest that accumulation of MBS RNA decay fragments does not always happen depending on the mRNA species and the model organisms used.

Temporal Pattern of cAMP Concentrations and α-Actin mRNA Expression in Skeletal Muscle of Cimaterol-Fed Rats

  • Kim, Y.S.;Duguies, M.V.;Kim, Y.H.;Vincent, D.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.5
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    • pp.528-533
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    • 1997
  • Twenty four female Sprague-Dawley rats weighing about 190 g were used to examine changes in muscle cAMP concentrations and steady-state levels of skeletal muscle ${\alpha}$-actin mRNA during chronic administration of cimaterol, a ${\beta}$-adrenergic agonist. Cimaterol was mixed in a powdered rat diet at 10 mg/kg diet. At 3 and 21 days after the start of treatment, skeletal muscle and heart samples were collected for the measurement of cAMP concentrations and skeletal muscle ${\alpha}$-actin mRNA levels. Cimaterol increased (p < 0.01) body weight gain gradually during the first seven days of the trial period, but not thereafter. Most skeletal muscle weights and the ratio of muscle weight to body weight were increased (p < 0.05) by cimaterol treatment both at 3 and 21 days. Heart weight was also increased (p < 0.05) by cimaterol treatment at 3 and 21 days, but the ratio of heart weight to body weight was increased (p < 0.05) only at 3 day. Cimaterol decreased (p < 0.05) cAMP concentration of gastrocnemius muscle at both 3 and 21 days after treatment. However, cimaterol tended (p = 0.07) to increase cAMP concentration at 3 days in the heart. Cimaterol tended (p = 0.08) to increase the steady-state level of ${\alpha}$-actin mRNA by 60% in gastrocnemius muscle at 3 days but had no effect at 21 days. The results indicate that the pattern of hypertrophic response to chronic dietary administration of cimaterol is different between cardiac and skeletal muscle. In skeletal muscles it appears that the hypertrophy induced by cimaterol is partly due to stimulated myofibrillar protein synthesis at a pre-translational level.

Cloning and mRNA Expression of an Actin cDNA from the Mulberry Longicorn Beetle, Apriona germari

  • Gui, Zhongzheng;Lee, Kwang Sik;Wei, Yadong;Yoon, Hyung Joo;Kim, Iksoo;Guo, Xijie;Sohn, Hung Dae;Jin, Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.2
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    • pp.187-191
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    • 2004
  • Actin is a ubiquitous and highly conserved protein found in eukaryotic organisms. In this study, we describe the cDNA cloning and mRNA expression of an actin gene from the mulberry longicorn beetle, Apriona germari. The A. germari actin cDNA is 1524 bp containing a complete 1128 bp open reading frame that encodes a polypeptide of 376 amino acid residues with a predicted molecular weight of about 41.5 kDa. The deduced amino acid sequence of the A.germari actin cDNA showed 99% protein sequence identity to Homalodisca coagulata actin, differing at only two amino acid positions, and 92-98% protein sequence identity to known insect species actins. The predicted three-dimensional structure of A. germari actin revealed the four residue hydrophobic pulg loop characteristic of the actin family. Northern blot analysis showed that A. germari actin is highly expressed in epidermis and muscle, and less strongly in midgut, but not in the fat body of A. germari larva.

RT- PCR Analysis of Vitellogenin Gene Expression in Bombina orientalis (무당개구리 비텔로제닌 유전자의 발현의 RT- PCR 검출법)

  • 계명찬;이명식;강희정;정경아;안혜선
    • Korean Journal of Environmental Biology
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    • v.22 no.2
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    • pp.329-335
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    • 2004
  • To develop a biomarker for the monitoring of the contamination of estrogenic endocrine disrupters in the aquatic environment, reverse transcription -polymerase chain reaction (RT-PCR) analysis of vitellogenin (Vg) mRNA expression was optimized in Bombina orientalis, a Korean red bellied toad species. Based on partial cDNA sequences of both Vg and beta actin genes of B. orientalis, specific primers for RT-PCR of Vg and beta actin mRNAs were developed. Semiquantitative RT-PCR of the Vg mRNA in liver was optimized using a beta actin mRNA as an internal control in both sexes. In female RT-PCR using $1\;\mu{g}$ of the liver cDNA resulted in a linear increment in the PCR product of Vg from 18 to 34 cycles of amplification. In male, on the contrary, the RT- PCR product was first detected at 30 cycles of amplification and a linear increment was observed from 30 to 40 cycles of amplification, suggesting that male B. orientalis expresses minute amount of Vg mRNA which is a $2^{-12}$ equivalent of female. In conclusion, the optimized protocol for semiquantitative RT-PCR analysis of Vg mRNA level in B. orientalis male liver will be useful for the environmental monitoring the xenoestrogen contamination in the freshwater environment in Korea.

Gene Expression upon Development of In Vitro Potato Microtuber (감자 기내 소괴경 발달에 따른 유전자 발현)

  • 홍주봉
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.309-321
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    • 1987
  • Differential gene expressions of patatin, proteinase inhibitor II, PAPI, rbcS and actin in potato microtuber have been examined. Microtubers from the several different stages of development were collected and their protein and mRNA patterns were examined. SDS-PAGE of microtuber proteins revealed that developmental changes in protein should be analogous to that of potatoes grown in the field. The level of patatin mRNA was the highest at the 30th day of development. Proteinse inhibitor IImRNA level was at the highest at the 15th day and decreased thereafter. The levels of PAPI mRNA, rbcS mRNA and actin mRNA were low throughout the time course examined.

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The Effect of Theophylline on Eotaxin mRNA Expression in Pulmonary Epithelial Cell Line A549 (인체 기도상피세포주에서 Theophylline에 의한 Eotaxin mRNA 발현억제)

  • Han, Min-Soo;Yoo, Jee-Hong;Kang, Hong-Mo
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.6
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    • pp.898-908
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    • 2000
  • Background : Eotaxin a CC chemokine specific for eosinophils, is implicated in the pathogenesis of asthma by recruiting eosinophils into the airways. Theophylline has been used for the treatment of asthma and recently was proposed to have an anti-inflammatory action. The aim of this study is to examine whether theophylline may inhibit the eosinophilic airway inflammation by reducing the expression of eotaxin. Methods : The expression of eotaxin mRNA was assessed by Northern analysis in A549 cells 4 h after stimulation with TNF-$\alpha$ or IL-1$\beta$. And then, theophylline was added to A549 cells stimulated with 0.1 ng/mL IL-1$\beta$. Results : Eotaxin mRNA expression rates induced by 0.1, 1, 10 ng/mL TNF-$\alpha$ as compared with $\beta$-actin, were 7%, 22%. 28%, respectively. Eotaxin mRNA expression rates induced by 0.01, 0.1, 1, 10 ng/mL IL-1$\beta$, as compared with $\beta$-actin, were 10%, 42%, 63%, 72%, respectively. Eotaxin mRNA expression rates after the addition of 0, 0.001, 0.01, 0.1 ${\mu}M$ dexamethasone induced by 10 ng/mL TNF-$\alpha$ as compared with $\beta$-actin, were 27%, 18%, 8%, respectively. Eotaxin mRNA expression rate after the addition of 0, 0.001, 0.01, 0.1 ${\mu}M$ dexamethasone induced by 0.1 ng/mL IL-1$\beta$ as compared with $\beta$-actin, were 43%, 47%, 12%, 8%, respectively. Eotaxin mRNA expression rates after the addition of 0, 0.001, 0.01, 0.1, 1, 10 mM theophylline induced by 0.1 ng/mL IL-1$\beta$, as compared with $\beta$-actin, were 48%, 40%, 33%, 22%, 16%, 14%, respectively. Conclusion : These results suggest that theophylline may reduce eosinophil infiltration of the airway at least in part by reducing the expression of eotaxin under the conditions of these experiments.

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Effect of ${\beta}-Estradiol$ on the Growth of Primary Rabbit Proximal Tubule Cells in Serum-free Medium (${\beta}-Estradiol$이 토끼 근위 세뇨관 상피세포의 성장에 미치는 영향)

  • Park, Sang-Ho;Chung, Joo-Ho;Ko, Kye-Chang;Jung, Jee-Chang
    • The Korean Journal of Pharmacology
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    • v.29 no.1
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    • pp.73-83
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    • 1993
  • In order to examine the effect of ${\beta}-estradiol$ on the cell growth, using a primary rabbit kidney poximal tubule cell culture system. We investigated the effect of ${\beta}-estradiol$ on alpha 1 (IV) collagen and ${\beta}-actin$ mRNA levels from primary rabbit kidney cell cultures, and also the effects of 3 growth factors and ${\beta}-estradiol$ supplementation on the growth of primary rabbit kidney proximal tubule cells in the serum-free medium. 1 nM of ${\beta}-estradiol$ showed a sizable potentiation effect on the growth of the proximal tubule cell in serum-free medium, but higher concentration (> 10 nM) of estradiol indeed inhibited the growth. In the absence of hydrocortisone as a growth supplement in serum-free medium, ${\beta}-estradiol$ caused to potentiate the growth of the cell. In the presence of hydrocortisone, ${\beta}-estradiol$ also potentiated the growth of the proximal tubule cells. According to the Northern analysis, ${\beta}-estradiol$ increased the level of ${\beta}-actin$ mRNA, although mRNA level of the alpha I(IV) collagen was not changed significantly.

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Neuronal function and dysfunction of CYFIP2: from actin dynamics to early infantile epileptic encephalopathy

  • Zhang, Yinhua;Lee, Yeunkum;Han, Kihoon
    • BMB Reports
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    • v.52 no.5
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    • pp.304-311
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    • 2019
  • The cytoplasmic FMR1-interacting protein family (CYFIP1 and CYFIP2) are evolutionarily conserved proteins originally identified as binding partners of the fragile X mental retardation protein (FMRP), a messenger RNA (mRNA)-binding protein whose loss causes the fragile X syndrome. Moreover, CYFIP is a key component of the heteropentameric WAVE regulatory complex (WRC), a critical regulator of neuronal actin dynamics. Therefore, CYFIP may play key roles in regulating both mRNA translation and actin polymerization, which are critically involved in proper neuronal development and function. Nevertheless, compared to CYFIP1, neuronal function and dysfunction of CYFIP2 remain largely unknown, possibly due to the relatively less well established association between CYFIP2 and brain disorders. Despite high amino acid sequence homology between CYFIP1 and CYFIP2, several in vitro and animal model studies have suggested that CYFIP2 has some unique neuronal functions distinct from those of CYFIP1. Furthermore, recent whole-exome sequencing studies identified de novo hot spot variants of CYFIP2 in patients with early infantile epileptic encephalopathy (EIEE), clearly implicating CYFIP2 dysfunction in neurological disorders. In this review, we highlight these recent investigations into the neuronal function and dysfunction of CYFIP2, and also discuss several key questions remaining about this intriguing neuronal protein.