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At least 19 recordsLinked to original sources

Acclimatization in a hot, humid environment: body fluid adjustments.

Four trained men worked 4 h/day at 40-50% of their maximum aerobic capacity first for 3 days at 25 degrees C db, 18 degrees C wb and then for 10 consecutive days at 45 degrees C db, 32 degrees C wb. Between days 1 and 2 of heat exposure mean total circulating protein (TCP) and plasma volume (PV) increased 11.6% and 9%, respectively. Preexposure TCP and PV increased until day 6 of heat exposure. Of the protein fractions beta-globulins underwent the largest relative increase. During work movement of protein into and out of the vascular compartment was similar in control and acclimatizing subjects but the latter generally maintained a greater amount of protein and fluid within the vascular volume. There was no evidence of salt and water retention. The increase in vascualr volume was ascribed to transfer of interstitial protein and water to the vascular volume. Regression coefficients indicated significant correlations for changes in plasma volume versus heart rate, stroke volume, and cardiac output during acclimatization. It was concluded that the most critical event in heat acclimatization is the expansion of the plasma volume.

Acclimatization

Homology of TcpN, a putative regulatory protein of Vibrio cholerae, to the AraC family of transcriptional activators.

The nucleotide sequence has been determined for the gene designated tcpN, encoding a putative regulatory protein within the tcp gene cluster associated with the biosynthesis and assembly of the toxin-coregulated pilus of Vibrio cholerae. It is preceded by a powerful transcriptional terminator which presumably delimits the major tcp operon, but at its 3' end is translationally coupled to the gene, tcpJ, encoding the TCP pilin signal peptidase. The tcpN gene encodes a putative 276-residue protein of 31,890 Da. This TcpN shows a high degree of homology to the transcriptional activators, Rns, associated with pilus biosynthesis in enterotoxigenic Escherichia coli, and to VirF, which controls the Yersinia virulence regulon. This homology also extends to the C termini of other members of the AraC family of transcriptional regulators, including RhaS, RhaR and CelD.

Amino Acid Sequence

Function in protein folding of TRiC, a cytosolic ring complex containing TCP-1 and structurally related subunits.

T-complex polypeptide 1 (TCP-1) was analyzed as a potential chaperonin (GroEL/Hsp60) equivalent of the eukaryotic cytosol. We found TCP-1 to be part of a hetero-oligomeric 970 kDa complex containing several structurally related subunits of 52-65 kDa. These members of a new protein family are assembled into a TCP-1 ring complex (TRiC) which resembles the GroEL double ring. The main function of TRiC appears to be in chaperoning monomeric protein folding: TRiC binds unfolded polypeptides, thereby preventing their aggregation, and mediates the ATP-dependent renaturation of unfolded firefly luciferase and tubulin. At least in vitro, TRiC appears to function independently of a small co-chaperonin protein such as GroES. Folding of luciferase is mediated by TRiC but not by GroEL/ES. This suggests that the range of substrate proteins interacting productively with TRiC may differ from that of GroEL. We propose that TRiC mediates the folding of cytosolic proteins by a mechanism distinct from that of the chaperonins in specific aspects.

Adenosine Triphosphatases

Functional Characterization of the Oat (Avena sativa L.) TCP Transcription Factor AsTCP38 Reveals Its Role in Low-Nitrogen Stress Tolerance.

Nitrogen limitation restricts plant growth, development, and yield in crops and forage species. Although TCP transcription factors are implicated in diverse abiotic-stress responses, the functions of most TCP genes in oat remain unclear. Here, we cloned and characterized the AsTCP38 gene, which is 1215 bp long and encodes a 405-amino-acid protein. The predicted protein contains a conserved TCP domain and shares its highest sequence similarity with Arabidopsis thaliana (A. thaliana) AtTCP15. The AsTCP38 protein localized to the nucleus, and promoter analysis identified cis-elements associated with light, hormone, and stress responses. We generated AsTCP38-overexpressing A. thaliana and wheat plants and screened an oat leaf yeast cDNA library for candidate interacting proteins. In these heterologous overexpression lines, AsTCP38 overexpression was associated with greater abscisic acid (ABA) sensitivity and improved seedling growth under low-nitrogen conditions. Changes in antioxidant-enzyme activities, nitrogen-metabolism-related enzyme activities, and endogenous hormone contents were also observed. Together, these findings suggest that AsTCP38 may participate in low-nitrogen responses and provide a basis for further functional studies in oat. Direct regulatory targets and the contribution of AsTCP38 to low-nitrogen adaptation in oat remain to be established.

Avena

Pleiotropic mutation in a tendril TCP gene underlies the yield-enhancing multiple-flowering trait in summer squash (Cucurbita pepo).

Crop yield is a focal point in plant breeding. Regulation of lateral budding through apical dominance was a central target of crop domestication, directly affecting crop production. The young fruits of Cucurbita pepo, summer squash, are produced on plants characterized by apical dominance and differentiation of a single flower bud per leaf axil. A single recessive mutation, mf, results in differentiation of more than one flower per leaf axil, thereby directly increasing production because of the continual day-to-day harvest of the summer squash crop. Positional cloning of the Cucurbita pepo mf (Cpmf) gene denoted a frameshift mutation in a TCP transcription factor, Cp4.1LG13g07780, as causative for the increase in axillary flowering. Cpmf is an ortholog of a tendril-development TCP gene in other cucurbits, and likewise, the recessive allele of Cpmf is associated with distorted tendril development. Gene function is context dependent, and we propose that multiple flowering is a unique pleiotropic attribute of mutation in a tendril-development gene of C. pepo. Characterization of a C. pepo collection confirmed a significant association of the Cpmf mutation with multiple flowering and showed that the mutant allele is absent in ancestral C. pepo and one of its two cultivated subspecies. The beneficial mutation occurred and was selected after the domestication of the other subspecies, during its cultivation for young fruit production. We demonstrate the discovery of a causative yield-increasing sequence variant and its practical utilization in breeding. Our findings provide a molecular target for creation of high-yielding, multiple-flowering summer squash cultivars through marker-assisted breeding or precise genome editing.

Cucurbita

CCT8 drives colorectal cancer progression via the RPL4-MDM2-p53 axis and immune modulation.

PURPOSE: Colorectal cancer (CRC) ranks high in global mortality, emphasizing the need for effective interventions. The aim of the research is to elucidate the oncogenic role of CCT8 in CRC and its interaction with RPL4 in the RPL4-MDM2-p53 axis. METHODS: TIMER 2.0, TCGA, and GTEx databases were used to analyze CCT8 expression patterns in CRC. Immunohistochemistry was performed to examine CCT8 distribution in CRC tissues and adjacent non-tumor tissues. Functional assays, including CCK-8, transwell, wound-healing, and flow cytometry, were conducted using DLD-1 and HCT116 cell lines to assess the effects of CCT8 on cell proliferation, migration, invasion, and apoptosis. Gene set enrichment analysis, protein-protein interaction network analysis, and co-immunoprecipitation were performed to explore the interaction between CCT8 and RPL4 and their role in the RPL4-MDM2-p53 pathway. Additionally, gene set variation analysis was applied to investigate the relationship between CCT8/RPL4 expression and immune infiltration patterns in CRC. RESULTS: CCT8 was significantly upregulated in CRC and associated with tumor progression. Mechanistically, CCT8 potentially synergizes with RPL4 concluded from their positive correlation and similar immune infiltration patterns, influencing the RPL4-MDM2-p53 axis and contributing to p53 ubiquitination and degradation. CONCLUSION: These findings underscore the oncogenic significance of CCT8 in CRC and shed light on its molecular mechanisms, paving the way for potential therapeutic applications.

Humans

A testis-expressed Zn finger gene (ZNF76) in human 6p21.3 centromeric to the MHC is closely linked to the human homolog of the t-complex gene tcp-11.

A novel testis-expressed Zn finger gene (ZNF76) was identified by screening cDNA libraries with cosmids derived from 6p21. ZNF76 is a member of the GLI-Krüppel family of DNA binding proteins. It is conserved in mouse where transcription in testis is initiated at Day 20 after birth. The mouse tcp-11 gene is located in the distal inversion of the t-complex and is developmentally regulated in the same manner as ZNF76. The human homolog of tcp-11 was isolated to allow a precise chromosomal localization. By using a combination of somatic cell hybrids, radiation hybrids, metaphase and interphase fluorescent in situ hybridization, and pulsed-field gel electrophoresis, we mapped the two genes to the 6p21.2 to 6p21.3 region and linked them to each other within 300 kb of DNA, approximately 2 Mb centromeric to the major histocompatibility complex.

Amino Acid Sequence

Exogenous gibberellin promotes lateral branch development in stumping Pinus yunnanensis by regulating endogenous hormones and TCP genes.

While plant stumping can break apical dominance, promote lateral branching, and thereby increase yield, a significant technical challenge remains in further enhancing the coppicing regrowth capacity after stumping. This study used one-year-old Pinus yunnanensis Franch seedlings as subjects. Through exogenous application of gibberellin 3 (GA3) and Paclobutrazol (PAC), it investigated the regulatory role of gibberellin (GA) in sprouting and identified TCP genes related to branching within the GA pathway. Results indicate that exogenous hormone treatments altered endogenous hormone levels in P. yunnanensis seedlings. Exogenous GA3 treatment significantly increased endogenous IPA, ABA, GA3, GA1, SA, and SAG content. Correlation analysis with sprout number revealed a positive relationship between endogenous hormones and sprout number, indicating that exogenous GA3 significantly influenced sprout number. After hormone application, the cumulative number of sprouts increased with seedling age across all treatments. Therefore, to further investigate whether changes in sprout number were caused by gene alterations in the GA signaling pathway, this study analyzed the GA signaling pathway and its associated transcription factors. Fifteen TCP family members were screened from the P. yunnanensis transcriptome, phylogenetically classified into Class I and Class II subfamilies. Conserved motif sites within the Class II family co-occurred, exhibiting high conservation. Genes from the Class II subfamily were cloned, and their differential expression across various tissues was analyzed via real-time quantitative PCR. Expression patterns under different hormone treatments were also examined. Our results reveal that PyTCP4, PyTCP6, and PyTCP20 are significantly upregulated by PAC (a GA synthesis inhibitor) and show strong negative correlations with sprouts number, while GA3 treatment partially alleviates this effect. These findings provide direct evidence that TCP transcription factors act as repressors of shoot branching, linking their hormone-responsive expression to the regulation of sprouts formation in P. yunnanensis. These results indicate that TCP transcription factors play a crucial role in shoot and root emergence, with their expression influenced by exogenous hormones, thereby affecting the growth and development of emerging shoots and roots. This provides a reference and basis for analyzing the expression patterns and functions of TCP genes in P. yunnanensis, facilitating further research into their functions.

Gibberellins

Characterization of specific binding sites for [3H]-1,3-di-o-tolyl-guanidine (DTG) in the rat glioma cell line C6-BU-1.

The aim of the present study was to find out if a cell line of glial origin possesses sigma and/or phencyclidine (PCP) binding sites. Binding of [3H]1,3-di-o-tolyl-guanidine (DTG), a highly selective ligand for sigma binding sites, and of [3H]N-[1-(2-thienyl)cyclohexyl] piperidine ([3H]TCP), a radioligand specific for PCP receptors, to C6-BU-1 glioma cells was investigated. Binding of [3H]DTG to C6-BU-1 cell membranes was reversible, saturable (Bmax = 10.5 pmol/mg protein), and of high affinity (KD = 26 nM). C6-BU-1 cells do not possess PCP receptors as indicated by negligible specific binding of [3H]TCP to C6-BU-1 cell membranes. Specific binding of [3H]DTG was reduced in the presence of Ca2+ and to a lesser extent by Mg2+. The rank order of potency of various PCP and sigma ligands was DTG > (+)3-[(3-hydroxy-phenyl)-N-n-propyl-piperidine] [(+)3-PPP] > haloperidol > pentazocine > (-)3-PPP > PCP > metaphit > dextromethorphan > (-)butaclamol > (+)butaclamol > (-)N-allylnormetazocine [(-)SKF 10,047] > MK801 > (+)SKF 10,047 > ketamine. The drug specificity, confirmed by a reversed stereoselectivity for the benzomorphan opiate SKF 10,047, indicated that these sites correspond to a subtype of sigma binding sites, the so-called sigma 2 binding site. Thus, the C6-BU-1 cell line is the first glial cell line demonstrated to have sigma 2 binding sites.

Animals

Effect of surfactants on antibiotic resistance.

The effectiveness of surfactants as potentiators of antibiotic activity on several resistant strains of bacteria, selected from clinical sources and laboratory collections, was studied using a tube dilution assay. Bacterial strains included members of the Enterobacteriaceae and staphylococci. Cetyltrimethylammonium bromide (CTAB), Tween 80 (Tw80), a mixture of n-alkyldimethyl betaines (L14), and alpha-(2,4,5-trichlorophenoxy) propionic acid (TCP) were tested in combination with pencillin G (PenG), methicillin (Met), streptomycin (Sm), polymyxin B (PmB), and chlortetracycline (CTC). Growth response to the drug combinations was compared with the response to each drug alone. CTAB and L14 but not Tw80 or TCP were found to potentiate the activity of CTC on strains of Escherichia coli, Proteus mirabilis, and Klebsiella pneumoniae. Studies on the inhibition of protein synthesis by CTC in cells of a strain of E. coli suggested that the surfactants increased the uptake of antibiotic into the cells. CTAB and L14 almost completely sensitized strains of P. mirabilis, Serratia marcescens, K. pneumoniae, and E. coli to PmB. With the exception of K. pneumoniae, TCP was also effective in potentiating the activity of PmB on the above strains whereas Tw80 showed potentiation only with a strain of E. coli. CTAB and L14 but not TCP or Tw80 potentiated the activity of PenG but not Met on strains of staphylococci. Studies of penicillinase in the cells suggested that the surfactants inhibited the formation of this enzyme possibly at the level of induction. None of the surfactants were found to potentiate the activity of Sm.

Anti-Bacterial Agents

Integrated analysis of ATAC-seq and RNA-seq reveals the TCP-ARF molecular module related to pathogenic process of phytoplasma infection in Paulownia fortunei.

BACKGROUND: Witches’ broom is an important disease of the Paulownia fortunei. Understanding the pathogenesis of witches’ broom is a prerequisite for its prevention and control. Phytoplasma is the pathogen of Paulownia witches’ broom. RESULTS: We investigated the changes in chromatin accessibility before and after phytoplasma infection in Paulownia fortunei by analyzing the DNA accessibility (ATAC-seq). In phytoplasma-infected P. fortunei (PFI) compared to healthy samples (PF), the closed regions of chromatin(1187 regions) were three times more than the open regions (352 regions). Fifty one percent of the accessible chromatin regions were overlapped with either H3K27ac or H3K9ac peaks. The closed regions were enriched in the conserved motif TGGGC[CT] that is recognized by the TCP transcription factor family. The closed regions in PFI are intersected with ARF family gene locus. The gene PfARF3 was verified to interact with the PfTCP23 transcription factor. The PfTCP23 was predicted to be interacted with the effector pawb44 in the pathogen of phytoplasma. CONCLUSIONS: The phytoplasma infection in P. fortunei is involved in the chromatin changes of the DNA accessibility and histone modification. The binding regions of TCP23 were found to be changed mostly in the accessibility between PFI and PF. The TCP-ARF module was found to be the possible regulatory module inducing the crinkled leaf trait.

RNA-Seq

TCP pilus biosynthesis in Vibrio cholerae O1: gene sequence of tcpC encoding an outer membrane lipoprotein.

The nucleotide sequence of the tcpC gene has been determined. It encodes a 53995-Da protein precursor with a signal sequence and cleavage site typical of a number of outer membrane lipoproteins, which are cleaved by the equivalent of signal peptidase II (Lsp) of Escherichia coli. The location of the tcpC gene is such that it is predicted to be translationally coupled to the 5' and 3' flanking genes, tcpY and tcpD, respectively, indicating that it forms part of an operon. Together with the lipoprotein signal sequence and the several hydrophobic domains it seems likely that TcpC is a surface-anchored trans-outer membrane lipoprotein.

Amino Acid Sequence

Thermoregulation of the pap operon: evidence for the involvement of RimJ, the N-terminal acetylase of ribosomal protein S5.

Our previous work showed that pap pilin gene transcription is subject to a thermoregulatory control mechanism under which pap pilin is not transcribed at a low temperature (23 degrees C) (L. B. Blyn, B. A. Braaten, C. A. White-Ziegler, D. H. Rolfson, and D. A. Low, EMBO J. 8:613-620, 1989). In order to isolate genes involved in this temperature regulation of gene expression, chromosomal mini-Tn10 (mTn10) mutations that allowed transcription of the pap pilin gene at 23 degrees C were identified, and the locus was designated tcp, for "thermoregulatory control of pap" (C. A. White-Ziegler, L. B. Blyn, B. A. Braaten, and D. A. Low, J. Bacteriol. 172:1775-1782, 1990). In the present study, quantitative analysis showed that the tcp mutations restore pap pilin transcription at 23 degrees C to levels similar to those measured at 37 degrees C. By in vivo recombination, the tcp mutations were mapped to phage E4H10S of the Kohara library of the Escherichia coli chromosome (Y. Kohara, K. Akiyama, and K. Isono, Cell 50:495-508, 1987). The tcp locus was cloned by complementation, in which a 1.3-kb DNA fragment, derived from the Kohara phage, was shown to restore thermoregulation to the mTn10 mutants. DNA sequencing revealed two open reading frames (ORFs) encoding proteins with calculated molecular masses of 22.7 and 20.3 kDa. The sequence of the 22.7-kDa ORF was identical to that of rimJ, the N-terminal acetylase of the ribosomal protein S5. The gene encoding the 20.3-kDa ORF, designated g20.3 here, did not display significant homology to any known DNA or protein sequence. On the basis of Northern (RNA) blot data, rimJ and g20.3 are located within the same operon. Two of the mTn10 transposons in the thermoregulatory mutants were inserted within the coding region of rimJ, indicating that the RimJ protein plays an important role in the temperature regulation of pap pilin gene transcription. However, rimJ itself is not thermoregulated, since rimJ transcripts were detected at both 23 and 37 degrees C. Disruption of the g20.3 gene by insertion and deletion mutagenesis did not affect thermoregulation of the pap operon, suggesting that, although g20.3 lies within the same operon as rimJ, it does not play a role in thermoregulation.

Acetylation

Distortion of transmission ratio by a candidate t complex responder locus transgene.

The mouse t complex responder locus (Tcr) is centrally involved in the phenomenon of male-specific transmission ratio distortion (TRD) through its action in haploid germ cells. Previously, we identified a candidate gene, Tcp-10b, whose t allele generates alternatively spliced transcripts. The full-length Tcp-10bt transcript is present in pre- and postmeiotic germ cells and encodes a product that is virtually identical with that encoded by the wild-type allele. The alternatively spliced t-specific transcript is observed in post-meiotic haploid spermatids and would encode an altered polypeptide that could convey the Tcrt phenotype. To assess their function, we have introduced constructs representing each Tcp-10bt transcript into transgenic mice. Breeding experiments demonstrate that these two constructs alter the transmission ratios of t haplotypes from male mice, but in opposite directions. The results provide support for the hypothesis that Tcp-10bt is a component of the Tcr locus.

Animals

NMDA receptor activation stimulates phospholipase A2 and somatostatin release from rat cortical neurons in primary cultures.

We have recently shown that glutamate exerts a stimulatory action on somatostatin secretion in cortical neurons essentially through NMDA receptor sites. Here, we investigated whether arachidonic acid release could be modified after NMDA receptor activation in cortical neurons in primary culture. We also studied whether pharmacological manipulation of phospholipase A2 could modify somatostatin release. We found that both glutamate and NMDA (N-methyl-D-aspartate) stimulated [3H]arachidonic acid release. NMDA-evoked arachidonic acid release was inhibited by MK-801 and TCP (two NMDA receptor-type antagonists), or by mepacrine, an inhibitor of phospholipase A2. NMDA-induced somatostatin release was inhibited by MK-801, mepacrine and by another phospholipase A2 inhibitor, p-bromophenacylbromide (pBPB). However, responses to NMDA were unaffected by H7, NDGA (nordihydroguaiaretic acid), indomethacin or by RHC 80267 (inhibitors of protein kinase C, lipooxygenase, cyclooxygenase and diacylglycerol lipase, respectively). Mepacrine (greater than or equal to 100 microM) decreased NMDA-stimulated phosphatidylinositol (PI) hydrolysis and at higher concentrations (250 microM) was also able to inhibit basal release whereas pBPB had no effect in the range of concentrations tested. Neomycin (which inhibits phosphatidylinositol metabolism by binding strongly and selectively to inositol phospholipids) reduced by 30% the NMDA-stimulated somatostatin release, although chronic treatment of neurons with the phorbol ester 12-myristate, 13-acetate (PMA) had no effect on this response. Melittin, an activator of phospholipase A2, was able to stimulate both arachidonic acid release and somatostatin secretion. High-performance liquid chromatography (HPLC) analysis of tritiated metabolites released from cortical neurons under basal or NMDA-stimulated conditions revealed that [3H]arachidonic acid was the only metabolite detectable. Furthermore, external addition of arachidonic acid increased somatostatin secretion. Our results show a correlation between the two parameters studied.

Animals

Enzyme-linked immunosorbent assay for determination of antibodies to Vibrio cholerae toxin-coregulated pili.

An ELISA for determination of antibodies to V. cholerae TCP was developed. Since purified TCP preparations contained detectable amounts of LPS (as shown by ELISA and immunoelectron microscopy with anti-LPS polyclonal serum), a capture ELISA was used. In this test the plate was coated with anti-TCP monoclonal antibody followed by incubation with TCP fimbriae. By this procedure no LPS bound to the solid phase as shown by the loss of reactivity with anti-LPS serum. The capture ELISA allowed sensitive and specific determination of TCP antibodies in sera of rabbits immunized with classical but not El Tor V. cholerae strains. There was good agreement between results in the TCP ELISA and reactivity with the TcpA band in immunoblot analyses when antisera raised against classical and El Tor vibrios were studied.

Animals