Purification and characterization of histone H1 from meiotic cells of a monocotyledonous plant, Lilium longiflorum

Genome ◽  
1994 ◽  
Vol 37 (5) ◽  
pp. 736-741 ◽  
Author(s):  
C. Daniel Riggs
Keyword(s):  
Lilium Longiflorum ◽  
Cell Types ◽  
Histone H1 ◽  
Cellular Protein ◽  
Vegetative Cells ◽  
Unbound Fraction ◽  
Total Cellular Protein ◽  
The Difference ◽  
Different Cell Types ◽  
Sulfuric Acid Solutions

To identify molecules involved in regulating meiotic chromatin structure, nuclear proteins from meiocytes of Lilium longiflorum were chromatographed on hydroxylapatite and the bound and unbound proteins were examined. An abundant nuclear protein was purified from the unbound fraction and by the following criteria was identified as a histone H1 molecule. The protein is soluble in acidic (perchloric and sulfuric acid) solutions, and its amino acid composition and the sequence of its amino terminus are similar to that of known histone H1s. An antiserum was produced against the protein to facilitate subsequent studies. Immunoblotting experiments demonstrated that histone H1 immunostaining declines in the developmental interval spanning the diplotene to tetrads stages. Concommitant with this decline is the appearance of several lower molecular mass, cross-reacting proteins. Neither the identity nor roles of these proteins is known. Immunoblotting experiments also demonstrate that, while the level of the protein is relatively constant in nuclei prepared from meiotic and vegetative cells, histone H1 is apparently enriched in total cellular extracts of meiotic cells compared with vegetative cells. This difference was found to be at least 16-fold. I conclude that in meiotic cells, histone H1 accounts for more of the total cellular protein than it does in vegetative cells. The difference in its relative abundance as a percent of the total cellular protein is probably in part due to differences in the ratio of nuclear to cytoplasmic volume in the different cell types, or the purging of sporophytic proteins from the cytoplasm of the meiocytes, or both.Key words: meiosis, histone H1, immunoblotting, meiotic purging.

scholarly journals Insulin and Analogue Effects on Protein Degradation in Different Cell Types

2000 ◽  
Vol 276 (15) ◽  
pp. 11552-11558 ◽  
Author(s):  
Janet Fawcett ◽  
Frederick G. Hamel ◽  
Robert G. Bennett ◽  
Zoltan Vajo ◽  
William C. Duckworth
Keyword(s):  
Protein Degradation ◽  
Hepg2 Cells ◽  
Cell Types ◽  
Major Effect ◽  
Cellular Protein ◽  
Experimental Conditions ◽  
L6 Cells ◽  
Different Cell Types

In adult animals, the major effect of insulin on protein turnover is inhibition of protein degradation. Cellular protein degradation is under the control of multiple systems, including lysosomes, proteasomes, calpains, and giant protease. Insulin has been shown to alter proteasome activityin vitroandin vivo. We examined the inhibition of protein degradation by insulin and insulin analogues (LysB28,ProB29-insulin (LysPro), AspB10-insulin (B10), and GluB4,GlnB16,PheB17-insulin (EQF)) in H4, HepG2, and L6 cells. These effects were compared with receptor binding. Protein degradation was examined by release of trichloroacetic acid-soluble radioactivity from cells previously labeled with [3H]leucine. Short- and intermediate-lived proteins were examined. H4 cells bound insulin with an EC50of 4.6 × 10−9m. LysPro was similar. The affinity of B10 was increased 2-fold; that of EQF decreased 15-fold. Protein degradation inhibition in H4 cells was highly sensitive to insulin (EC50= 4.2 × 10−11and 1.6 × 10−10m, short- and intermediate-lived protein degradation, respectively) and analogues. Despite similar binding, LysPro was 11- to 18-fold more potent than insulin at inhibiting protein degradation. Conversely, although EQF showed lower binding to H4 cells than insulin, its action was similar. The relative binding potencies of analogues in HepG2 cells were similar to those in H4 cells. Examination of protein degradation showed insulin, LysPro, and B10 were equivalent while EQF was less potent. L6 cells showed no difference in the binding of the analogues compared with insulin, but their effect on protein degradation was similar to that seen in HepG2 cells except B10 inhibited intermediate-lived protein degradation better than insulin. These studies illustrate the complexities of cellular protein degradation and the effects of insulin. The effect of insulin and analogues on protein degradation vary significantly in different cell types and with different experimental conditions. The differences seen in the action of the analogues cannot be attributed to binding differences. Post-receptor mechanisms, including intracellular processing and degradation, must be considered.

scholarly journals The Heterocyst-Specific NsiR1 Small RNA Is an Early Marker of Cell Differentiation in Cyanobacterial Filaments

mBio ◽  
2014 ◽  
Vol 5 (3) ◽  
Author(s):  
Alicia M. Muro-Pastor
Keyword(s):  
Small Rna ◽  
Single Cells ◽  
Nitrogen Deficiency ◽  
Cell Types ◽  
Nitrogen Fixing ◽  
Vegetative Cells ◽  
Early Marker ◽  
Multicellular Organisms ◽  
Pcc 7120 ◽  
Different Cell Types

ABSTRACT Differentiation of single cells along filaments of cyanobacteria constitutes one of the simplest developmental patterns in nature. In response to nitrogen deficiency, certain cells located in a semiregular pattern along filaments differentiate into specialized nitrogen-fixing cells called heterocysts. The process involves the sequential activation of many genes whose expression takes place, either exclusively or at least more strongly, in those cells undergoing differentiation. In the model cyanobacterium Anabaena (Nostoc) sp. strain PCC 7120, increased transcription of hetR, considered the earliest detectable heterocyst-specific transcript, has been reported to occur in pairs or even in clusters of cells, thus making it difficult to identify prospective heterocysts during the early stages of differentiation, before any morphological change is detectable. The promoter of nsiR1 (nitrogen stress inducible RNA1), a heterocyst-specific small RNA, constitutes a minimal sequence promoting heterocyst-specific transcription. Using confocal fluorescence microscopy, I have analyzed expression of a gfp reporter transcriptionally fused to P nsiR1 . The combined analysis of green fluorescence (reporting transcriptional activity from P nsiR1 ) and red fluorescence (an indication of progress in the differentiation of individual cells) shows that expression of P nsiR1 takes place in single cells located in a semiregular pattern before any other morphological or fluorescence signature of differentiation can be observed, thus providing an early marker for cells undergoing differentiation. IMPORTANCE Cyanobacterial filaments containing heterocysts constitute an example of bacterial division of labor. When using atmospheric nitrogen, these filaments behave as multicellular organisms in which two different cell types (vegetative cells and nitrogen-fixing heterocysts) coexist and cooperate to achieve growth of the filament as a whole. The molecular basis governing the differentiation of individual vegetative cells, and thus the establishment of a one-dimensional pattern from cells that are apparently the same, remains one of the most intriguing aspects of this differentiation process. Recent evidence suggests that, at any given time, some cells in the filaments are more likely than others to become heterocysts when nitrogen limitation is encountered. The robust heterocyst-specific nsiR1 promoter, which is induced very early during differentiation, provides a valuable tool to analyze issues such as early candidacy or the possible role of transcriptional noise in determining the fate of specific cells in cyanobacterial filaments.

scholarly journals Differences in the intrinsic spatial dynamics of the chromatin contribute to cell differentiation

2019 ◽  
Vol 48 (3) ◽  
pp. 1131-1145
Author(s):  
She Zhang ◽  
Fangyuan Chen ◽  
Ivet Bahar
Keyword(s):  
Cell Differentiation ◽  
Cell Lines ◽  
Spatial Dynamics ◽  
Cell Types ◽  
Global Scale ◽  
Elastic Network Model ◽  
Chromosome Conformation ◽  
Highly Expressed Genes ◽  
The Difference ◽  
Different Cell Types

Abstract Advances in chromosome conformation capture techniques as well as computational characterization of genomic loci structural dynamics open new opportunities for exploring the mechanistic aspects of genome-scale differences across different cell types. We examined here the dynamic basis of variabilities between different cell types by investigating their chromatin mobility profiles inferred from Hi-C data using an elastic network model representation of the chromatin. Our comparative analysis of sixteen cell lines reveals close similarities between chromosomal dynamics across different cell lines on a global scale, but notable cell-specific variations emerge in the detailed spatial mobilities of genomic loci. Closer examination reveals that the differences in spatial dynamics mainly originate from the difference in the frequencies of their intrinsically accessible modes of motion. Thus, even though the chromosomes of different types of cells have access to similar modes of collective movements, not all modes are deployed by all cells, such that the effective mobilities and cross-correlations of genomic loci are cell-type-specific. Comparison with RNA-seq expression data reveals a strong overlap between highly expressed genes and those distinguished by high mobilities in the present study, in support of the role of the intrinsic spatial dynamics of chromatin as a determinant of cell differentiation.

scholarly journals Influence of Rearrangement of Actin Cytoskeleton on the Overall Material Properties of ATDC5 Cells During Chondrogenesis

2004 ◽  
Author(s):  
Madhavi Ayyalasomayajula ◽  
Sydha Salihu ◽  
Vincent Kish ◽  
Nilay Mukherjee
Keyword(s):  
Finite Element ◽  
Material Properties ◽  
Transverse Direction ◽  
Cell Types ◽  
Stress Fiber ◽  
Finite Element Models ◽  
Agarose Gel ◽  
Actin Structure ◽  
The Difference ◽  
Different Cell Types

ATDC5 cells, a prechondrocytic cell line, were cultured in 2% agarose gel-cell constructs. A major reorientation of actin structure (stress-fiber to punctate) was observed during the chondrogenesis of these cells. 2D finite element models of cell (considered as a composite with embedded actin fibers) in a 500μ3 gel were created for days 1 (prechondrocytes), 5, 8 & 12 (chondrocytes) of culture. The response of the cells to uniaxial compression of the gel-cell construct was studied. The model predicted that, as a result of rearrangement of actin, the cells became more compliant and less anisotropic. When the gel size was reduced to 20μ3, RUTL (Ratio of radial deformations of cell in the Transverse direction to Loaded direction) became significantly different between prechondrocytes and chondrocytes. The difference in force required to compress a 20μ3 gel-cell construct containing a prechondrocyte vs a chondrocyte to 15% deformation was 103nN (152.3 vs 48.8 nN). This information can be used to build MEMS based devices that can mechanically distinguish between the different cell types.

Induction of cell-specific ribosomal proteins in aggregation-competent nonmorphogenetic Dictyostelium discoideum

1990 ◽  
Vol 68 (11) ◽  
pp. 1281-1287 ◽  
Author(s):  
S. Ramagopal
Keyword(s):  
Ribosomal Proteins ◽  
Cell Types ◽  
Growth Cycle ◽  
Logarithmic Phase ◽  
Cellular Slime Mold ◽  
Developmentally Regulated ◽  
Vegetative Cells ◽  
Competent Cells ◽  
Cellular Slime ◽  
Different Cell Types

Vegetatively growing amoebae, if shaken in a starvation (nonnutrient) buffer, acquire aggregation competence, but do not embark on a morphogenetic program. The quantitative variation of ribosomal proteins in vegetative and aggregation-competent cells was compared by labeling the different cell types with [35S]methionine. Vegetative cells were examined at various phases of the growth cycle. No changes could be detected in the content of ribosomes or the apparent stoichiometry of ribosomal proteins in growing cells. In stationary phase cells, the net ribosome content declined to 15% of that observed in logarithmic phase, but the relative amounts of individual ribosomal proteins were not altered. Although aggregation-competent cells contained 30% less ribosomes compared with logarithmic phase cells, the total fraction of newly made ribosomal proteins was the same in both. In contrast to vegetative cells, distinct changes were induced in the ribosomal proteins of aggregation-competent cells. The composition of ribosomes in aggregation-competent phase resembled in every respect that observed in spore cells. As reported earlier, changes were found in all 12 of the developmentally regulated ribosomal proteins. For the majority of newly made ribosomal proteins during aggregation competence, the stoichiometry was similar to that in logarithmically growing cells. However, the relative synthesis of some was particularly higher (13- to 46-fold for A and L; 3- to 8-fold for D, E, S24, L3, S6, and L4) compared with logarithmic phase cells. About 18 proteins, which included the cell-specific ribosomal proteins L18, S10, S14, S16, and L11, were synthesized in lesser amounts than in logarithmic phase cells. It is concluded that the attainment of aggregation competence is sufficient for the induction of spore cell specific ribosomal proteins in Dictyostelium discoideum.Key words: cellular slime mold, ribosomal proteins, development.

scholarly journals Anaphase A Chromosome Movement and Poleward Spindle Microtubule Flux Occur At Similar Rates in Xenopus Extract Spindles

1998 ◽  
Vol 141 (3) ◽  
pp. 703-713 ◽  
Author(s):  
Arshad Desai ◽  
Paul S. Maddox ◽  
Timothy J. Mitchison ◽  
E.D. Salmon
Keyword(s):  
Somatic Cells ◽  
Cell Types ◽  
Chromosome Movement ◽  
Microtubule Depolymerization ◽  
Spindle Microtubule ◽  
Egg Extracts ◽  
Xenopus Egg ◽  
The Difference ◽  
Xenopus Egg Extracts ◽  
Different Cell Types

We have used local fluorescence photoactivation to mark the lattice of spindle microtubules during anaphase A in Xenopus extract spindles. We find that both poleward spindle microtubule flux and anaphase A chromosome movement occur at similar rates (∼2 μm/min). This result suggests that poleward microtubule flux, coupled to microtubule depolymerization near the spindle poles, is the predominant mechanism for anaphase A in Xenopus egg extracts. In contrast, in vertebrate somatic cells a “Pacman” kinetochore mechanism, coupled to microtubule depolymerization near the kinetochore, predominates during anaphase A. Consistent with the conclusion from fluorescence photoactivation analysis, both anaphase A chromosome movement and poleward spindle microtubule flux respond similarly to pharmacological perturbations in Xenopus extracts. Furthermore, the pharmacological profile of anaphase A in Xenopus extracts differs from the previously established profile for anaphase A in vertebrate somatic cells. The difference between these profiles is consistent with poleward microtubule flux playing the predominant role in anaphase chromosome movement in Xenopus extracts, but not in vertebrate somatic cells. We discuss the possible biological implications of the existence of two distinct anaphase A mechanisms and their differential contributions to poleward chromosome movement in different cell types.

Study of the Molecular Architecture of Keratin Filaments by Electron Microscopy

1982 ◽  
Vol 40 ◽  
pp. 118-119
Author(s):  
U. Aebi ◽  
P. Rew ◽  
T.-T. Sun
Keyword(s):  
Electron Microscopy ◽  
Structural Organization ◽  
Cell Types ◽  
Structural Features ◽  
Molecular Architecture ◽  
The Past ◽  
Keratin Filaments ◽  
Different Types ◽  
10 Nm Filaments ◽  
Different Cell Types

Various types of intermediate-sized (10-nm) filaments have been found and described in many different cell types during the past few years. Despite the differences in the chemical composition among the different types of filaments, they all yield common structural features: they are usually up to several microns long and have a diameter of 7 to 10 nm; there is evidence that they are made of several 2 to 3.5 nm wide protofilaments which are helically wound around each other; the secondary structure of the polypeptides constituting the filaments is rich in ∞-helix. However a detailed description of their structural organization is lacking to date.

Processing and Characterization of Recombinant von Willebrand Factor Expressed in Different Cell Types Using a Vaccinia Virus Vector

1992 ◽  
Vol 67 (01) ◽  
pp. 154-160 ◽  
Author(s):  
P Meulien ◽  
M Nishino ◽  
C Mazurier ◽  
K Dott ◽  
G Piétu ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Von Willebrand Factor ◽  
Human Fibroblasts ◽  
Active Form ◽  
Cell Types ◽  
Biologically Active ◽  
L Cells ◽  
Von Willebrand ◽  
Different Cell Types ◽  
Willebrand Factor

SummaryThe cloning of the cDNA encoding von Willebrand factor (vWF) has revealed that it is synthesized as a large precursor (pre-pro-vWF) molecule and it is now clear that the prosequence or vWAgll is responsible for the intracellular multimerization of vWF. We have cloned the complete vWF cDNA and expressed it using a recombinant vaccinia virus as vector. We have characterized the structure and function of the recombinant vWF (rvWF) secreted from five different cell types: baby hamster kidney (BHK), Chinese hamster ovary (CHO), human fibroblasts (143B), mouse fibroblasts (L) and primary embryonic chicken cells. Forty-eight hours after infection, the quantity of vWF antigen found in the cell supernatant varied from 3 to 12 U/dl depending on the cell type. By SDS-agarose gel electrophoresis, the percentage of high molecular weight forms of vWF varied from 39 to 49% relative to normal plasma for BHK, CHO, 143B and chicken cells but was less than 10% for L cells. In all cell types, the two anodic subbands of each multimer were missing. The two cathodic subbands were easily detected only in BHK and L cells. By SDS-PAGE of reduced samples, pro-vWF was present in similar quantity to the fully processed vWF subunit in L cells, present in moderate amounts in BHK and CHO and in very low amounts in 143B and chicken cells. rvWF from all cells bound to collagen and to platelets in the presence of ristocetin, the latter showing a high correlation between binding efficiency and degree of multimerization. rvWF from all cells was also shown to bind to purified FVIII and in this case binding appeared to be independent of the degree of multimerization. We conclude that whereas vWF is naturally synthesized only by endothelial cells and megakaryocytes, it can be expressed in a biologically active form from various other cell types.

Role of Transcriptional Read-Through in PRE Activity in Drosophila melanogaster

Acta Naturae ◽  
2016 ◽  
Vol 8 (2) ◽  
pp. 79-86 ◽  
Author(s):  
P. V. Elizar’ev ◽  
D. V. Lomaev ◽  
D. A. Chetverina ◽  
P. G. Georgiev ◽  
M. M. Erokhin
Keyword(s):  
Dna Sequences ◽  
Cell Types ◽  
Transcription Terminator ◽  
Response Elements ◽  
Polycomb Response Elements ◽  
The Individual ◽  
Multicellular Organisms ◽  
Different Cell Types ◽  
Main Factor

Maintenance of the individual patterns of gene expression in different cell types is required for the differentiation and development of multicellular organisms. Expression of many genes is controlled by Polycomb (PcG) and Trithorax (TrxG) group proteins that act through association with chromatin. PcG/TrxG are assembled on the DNA sequences termed PREs (Polycomb Response Elements), the activity of which can be modulated and switched from repression to activation. In this study, we analyzed the influence of transcriptional read-through on PRE activity switch mediated by the yeast activator GAL4. We show that a transcription terminator inserted between the promoter and PRE doesnt prevent switching of PRE activity from repression to activation. We demonstrate that, independently of PRE orientation, high levels of transcription fail to dislodge PcG/TrxG proteins from PRE in the absence of a terminator. Thus, transcription is not the main factor required for PRE activity switch.

MAPK: A Key Player in the Development and Progression of Stroke

2020 ◽  
Vol 19 (4) ◽  
pp. 248-256
Author(s):  
Yangmin Zheng ◽  
Ziping Han ◽  
Haiping Zhao ◽  
Yumin Luo
Keyword(s):  
Ischemic Stroke ◽  
Signaling Pathway ◽  
Complex Disease ◽  
Therapeutic Targets ◽  
Cell Types ◽  
Mapk Signaling ◽  
Mapk Signaling Pathway ◽  
Effective Prevention ◽  
Prevention And Treatment ◽  
Different Cell Types

Conclusion: Stroke is a complex disease caused by genetic and environmental factors, and its etiological mechanism has not been fully clarified yet, which brings great challenges to its effective prevention and treatment. MAPK signaling pathway regulates gene expression of eukaryotic cells and basic cellular processes such as cell proliferation, differentiation, migration, metabolism and apoptosis, which are considered as therapeutic targets for many diseases. Up to now, mounting evidence has shown that MAPK signaling pathway is involved in the pathogenesis and development of ischemic stroke. However, the upstream kinase and downstream kinase of MAPK signaling pathway are complex and the influencing factors are numerous, the exact role of MAPK signaling pathway in the pathogenesis of ischemic stroke has not been fully elucidated. MAPK signaling molecules in different cell types in the brain respond variously after stroke injury, therefore, the present review article is committed to summarizing the pathological process of different cell types participating in stroke, discussed the mechanism of MAPK participating in stroke. We further elucidated that MAPK signaling pathway molecules can be used as therapeutic targets for stroke, thus promoting the prevention and treatment of stroke.

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