scholarly journals Weaver Syndrome‐Associated EZH2 Protein Variants Show Impaired Histone Methyltransferase Function In Vitro

2016 ◽  
Vol 37 (3) ◽  
pp. 301-307 ◽  
Author(s):  
Ana S.A. Cohen ◽  
Damian B. Yap ◽  
M.E. Suzanne Lewis ◽  
Chieko Chijiwa ◽  
Maria A. Ramos‐Arroyo ◽  
...  
Keyword(s):  
Histone Methyltransferase ◽  
Weaver Syndrome ◽  
Protein Variants

scholarly journals Histone methyltransferase WHSC1 inhibits colorectal cancer cell apoptosis via targeting anti-apoptotic BCL2

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Yu Wang ◽  
Liming Zhu ◽  
Mei Guo ◽  
Gang Sun ◽  
Kun Zhou ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cancer Cell ◽  
Cell Apoptosis ◽  
B Cell Lymphoma ◽  
Histone Methyltransferase ◽  
Chemotherapeutic Agents ◽  
Cancer Cell Apoptosis ◽  
Active Transcription

AbstractWHSC1 is a histone methyltransferase that facilitates histone H3 lysine 36 dimethylation (H3K36me2), which is a permissive mark associated with active transcription. In this study, we revealed how WHSC1 regulates tumorigenesis and chemosensitivity of colorectal cancer (CRC). Our data showed that WHSC1 as well as H3K36me2 were highly expressed in clinical CRC samples, and high WHSC1 expression is associated with poorer prognosis in CRC patients. WHSC1 reduction promoted colon cancer cell apoptosis both in vivo and in vitro. We found that B cell lymphoma-2 (BCL2) expression, an anti-apoptotic protein, is markedly decreased in after WHSC1 depletion. Mechanistic characterization indicated that WHSC1 directly binds to the promoter region of BCL2 gene and regulate its H3K36 dimethylation level. What’s more, our study indicated that WHSC1 depletion promotes chemosensitivity in CRC cells. Together, our results suggested that WHSC1 and H3K36me2 modification might be optimal therapeutic targets to disrupt CRC progression and WHSC1-targeted therapy might potentially overcome the resistance of chemotherapeutic agents.

In-vitro Selection of Highly Stabilized Protein Variants with Optimized Surface

2001 ◽  
Vol 309 (3) ◽  
pp. 717-726 ◽  
Author(s):  
Andreas Martin ◽  
Volker Sieber ◽  
Franz X. Schmid
Keyword(s):  
In Vitro Selection ◽  
Protein Variants ◽  
Selection Of

scholarly journals In Vitro Histone Methyltransferase Assay

2008 ◽  
Vol 2008 (3) ◽  
pp. pdb.prot4939-pdb.prot4939 ◽  
Author(s):  
I. M. Fingerman ◽  
H.-N. Du ◽  
S. D. Briggs
Keyword(s):  
Histone Methyltransferase

The Histone Methyltransferase EZH2 Adopts a Nuclear Localization During Cleavage in Porcine Embryos Fertilized In Vitro.

2010 ◽  
Vol 83 (Suppl_1) ◽  
pp. 304-304
Author(s):  
Celeste E. Morrison ◽  
K. Park ◽  
X. Wang ◽  
R. A. Cabot
Keyword(s):  
Nuclear Localization ◽  
Histone Methyltransferase

Estimation of mutation rates based on the analysis of polypeptide constituents of cultured human lymphoblastoid cells.

Genetics ◽  
1988 ◽  
Vol 119 (3) ◽  
pp. 693-703
Author(s):  
E H Chu ◽  
M Boehnke ◽  
S M Hanash ◽  
R D Kuick ◽  
B J Lamb ◽  
...  
Keyword(s):  
Structural Gene ◽  
Somatic Mutations ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gene Mutations ◽  
Apparent Molecular Weight ◽  
Cell Generation ◽  
Mutation Rates ◽  
Cell Clones ◽  
Protein Variants

Abstract A subclone of a human diploid lymphoblastoid cell line, TK-6, with consistently high cloning efficiency has been used to estimate the rates of somatic mutations on the basis of protein variation detected by two-dimensional polyacrylamide gel electrophoresis. A panel of 267 polypeptide spots per gel was screened, representing the products of approximately 263 unselected loci. The rate of human somatic mutation in vitro was estimated by measuring the proportion of protein variants among cell clones isolated at various times during continuous exponential growth of a TK-6 cell population. Three mutants of spontaneous origin were observed, giving an estimated spontaneous rate of 6 x 10(-8) electrophoretic mutations per allele per cell generation (i.e., 1.2 x 10(-7) per locus per cell generation). Following treatment of cells with N-ethyl-N-nitrosourea, a total of 74 confirmed variants at 54 loci were identified among 1143 clones analyzed (approximately 601,000 allele tests). The induced variants include 65 electromorphs which exhibit altered isoelectric charge and/or apparent molecular weight and nine nullimorphs for each of which a gene product was not detected at its usual location on the gel. The induced frequency for these 65 structural gene mutants is 1.1 x 10(-4) per allele. An excess of structural gene mutations at ten known polymorphic loci and repeat mutations at these and other loci suggest nonrandomness of mutation in human somatic cells. Nullimorphs occurring at three heterozygous loci in TK-6 cells may be caused by genetic processes other than structural gene mutation.

scholarly journals Targeting Human Thrombus by Liposomes Modified with Anti-Fibrin Protein Binders

Pharmaceutics ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 642 ◽  
Author(s):  
Hana Petroková ◽  
Josef Mašek ◽  
Milan Kuchař ◽  
Andrea Vítečková Wünschová ◽  
Jana Štikarová ◽  
...  
Keyword(s):  
Amino Acid ◽  
Large Scale ◽  
Enzyme Linked Immunosorbent Assay ◽  
Ribosome Display ◽  
Amino Acid Peptide ◽  
His Tag ◽  
Insoluble Form ◽  
Protein Binders ◽  
Protein Variants

Development of tools for direct thrombus imaging represents a key step for diagnosis and treatment of stroke. Nanoliposomal carriers of contrast agents and thrombolytics can be functionalized to target blood thrombi by small protein binders with selectivity for fibrin domains uniquely formed on insoluble fibrin. We employed a highly complex combinatorial library derived from scaffold of 46 amino acid albumin-binding domain (ABD) of streptococcal protein G, and ribosome display, to identify variants recognizing fibrin cloth in human thrombus. We constructed a recombinant target as a stretch of three identical fibrin fragments of 16 amino acid peptide of the Bβ chain fused to TolA protein. Ribosome display selection followed by large-scale Enzyme-Linked ImmunoSorbent Assay (ELISA) screening provided four protein variants preferentially binding to insoluble form of human fibrin. The most specific binder variant D7 was further modified by C-terminal FLAG/His-Tag or double His-tag for the attachment onto the surface of nanoliposomes via metallochelating bond. D7-His-nanoliposomes were tested using in vitro flow model of coronary artery and their binding to fibrin fibers was demonstrated by confocal and electron microscopy. Thus, we present here the concept of fibrin-targeted binders as a platform for functionalization of nanoliposomes in the development of advanced imaging tools and future theranostics.

scholarly journals Transcriptional Repression by the Retinoblastoma Protein through the Recruitment of a Histone Methyltransferase

2001 ◽  
Vol 21 (19) ◽  
pp. 6484-6494 ◽  
Author(s):  
Laurence Vandel ◽  
Estelle Nicolas ◽  
Olivier Vaute ◽  
Roger Ferreira ◽  
Slimane Ait-Si-Ali ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Transcriptional Repression ◽  
Histone Deacetylases ◽  
Histone H3 ◽  
Histone Methyltransferase ◽  
S Phase ◽  
E2f Transcription Factor ◽  
Cycle Dependent

ABSTRACT The E2F transcription factor controls the cell cycle-dependent expression of many S-phase-specific genes. Transcriptional repression of these genes in G0 and at the beginning of G1by the retinoblasma protein Rb is crucial for the proper control of cell proliferation. Rb has been proposed to function, at least in part, through the recruitment of histone deacetylases. However, recent results indicate that other chromatin-modifying enzymes are likely to be involved. Here, we show that Rb also interacts with a histone methyltransferase, which specifically methylates K9 of histone H3. The results of coimmunoprecipitation experiments of endogenous or transfected proteins indicate that this histone methyltransferase is the recently described heterochromatin-associated protein Suv39H1. Interestingly, phosphorylation of Rb in vitro as well as in vivo abolished the Rb-Suv39H1 interaction. We also found that Suv39H1 and Rb cooperate to repress E2F activity and that Suv39H1 could be recruited to E2F1 through its interaction with Rb. Taken together, these data indicate that Suv39H1 is involved in transcriptional repression by Rb and suggest an unexpected link between E2F regulation and heterochromatin.

Human SP-A protein variants derived from one or both genes stimulate TNF-α production in the THP-1 cell line

2000 ◽  
Vol 278 (5) ◽  
pp. L946-L954 ◽  
Author(s):  
Guirong Wang ◽  
David S. Phelps ◽  
Todd M. Umstead ◽  
Joanna Floros
Keyword(s):  
Tumor Necrosis ◽  
Single Gene ◽  
Surfactant Protein ◽  
Functional Genes ◽  
Gene Products ◽  
Functional Differences ◽  
Tnf Α ◽  
Protein Variants ◽  
Necrosis Factor

In humans, two functional genes of surfactant protein (SP) A, SP-A1 and SP-A2, and several alleles of each functional gene have been characterized. SP-A is a multimeric molecule consisting of six trimers. Each trimer contains two SP-A1 molecules and one SP-A2 molecule. Until now, it has been unclear whether a single SP-Agene product is functional or whether there are functional differences either among alleles or between single-gene SP-A products and SP-A products derived from both genes. We tested the ability of in vitro expressed SP-A variants to stimulate tumor necrosis factor (TNF)-α production by THP-1 cells. We observed that 1) single-gene products and products derived from both genes stimulate TNF-α production, 2) there are differences among SP-A1 and SP-A2 alleles in their ability to stimulate TNF-α production, and 3) the increases in TNF-α production are lower after treatment with the SP-A1 alleles than after treatment with the SP-A2 alleles. Furthermore, coexpressed SP-As from SP-A1 and SP-A2 genes have a higher activity compared with SP-As from individual alleles or mixed SP-As from SP-A1and SP-A2 genes. These data suggest that the SP-A-induced increases in TNF-α levels differ among SP-A variants and appear to be affected by SP-A genotype and whether SP-A is derived from one or both genes.

scholarly journals Comparison of the ability of mammalian eEF1A1 and its oncogenic variant eEF1A2 to interact with actin and calmodulin

2017 ◽  
Vol 398 (1) ◽  
pp. 113-124 ◽  
Author(s):  
Oleksandra Novosylna ◽  
Annette Doyle ◽  
Dmytro Vlasenko ◽  
Mark Murphy ◽  
Boris Negrutskii ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Elongation Factor ◽  
Translation Elongation Factor ◽  
Translation Elongation ◽  
Oncogenic Potential ◽  
Regulatory Systems ◽  
Protein Variants ◽  
Homologous Tissue ◽  
Trna Binding

Abstract The question as to why a protein exerts oncogenic properties is answered mainly by well-established ideas that these proteins interfere with cellular signaling pathways. However, the knowledge about structural and functional peculiarities of the oncoproteins causing these effects is far from comprehensive. The 97.5% homologous tissue-specific A1 and A2 isoforms of mammalian translation elongation factor eEF1A represent an interesting model to study a difference between protein variants of a family that differ in oncogenic potential. We propose that the different oncogenic impact of A1 and A2 might be explained by differences in their ability to communicate with their respective cellular partners. Here we probed this hypothesis by studying the interaction of eEF1A with two known partners – calmodulin and actin. Indeed, an inability of the A2 isoform to interact with calmodulin is shown, while calmodulin is capable of binding A1 and interferes with its tRNA-binding and actin-bundling activities in vitro. Both A1 and A2 variants revealed actin-bundling activity; however, the form of bundles formed in the presence of A1 or A2 was distinctly different. Thus, a potential inability of A2 to be controlled by Ca2+-mediated regulatory systems is revealed.

Effects of 27 CYP3A4 Protein Variants on saxagliptin metabolism in vitro

2021 ◽  
Author(s):  
Qian liu ◽  
Qiu‐Geng Ou‐Yang ◽  
Qian‐Meng Lin ◽  
Xiang‐Ran Lu ◽  
Ya‐Qing Ma ◽  
...  
Keyword(s):  
Protein Variants
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