scholarly journals Analysis of the ratio of α- to β-globin and globin messenger ribonucleic acid content of fractionated rabbit erythroid bone-marrow cells

1979 ◽  
Vol 179 (3) ◽  
pp. 525-535 ◽  
Author(s):  
V A Mezl ◽  
E S Kawasaki ◽  
J A Hunt
Keyword(s):  
Bone Marrow Cells ◽  
Beta Globin ◽  
Globin Mrna ◽  
Constant Amount ◽  
Messenger Ribonucleic Acid ◽  
Mrna Ratio ◽  
Linear Reaction ◽  
Mrna Content ◽  
Cdna Hybridization ◽  
Better Than

The ratio of alpha- to beta-globin mRNA was measured by hybridization of a constant amount of highly purified alpha- or beta-globin cDNA (complementary DNA) with increasing amounts of RNA in the range up to 20% cDNA hybridization, where an essentially linear reaction is obtained. Statistical analysis indicates that the ratio of alpha- to beta-globin can be measured within a maximal error of +/- 0.3 and in most cases is better than +/- 0.15. Under these conditions there is no significant deviation from the ratio of 1.3 in the alpha- to beta-globin mRNA ratio of RNA isolated from erythroid cells rich in pronormoblasts through to reticulocytes. If the ratio of alpha- to beta-globin mRNA exceeded 1.7 or was less than 0.9 in pronormoblasts, it would be detected in these experiments. The overall globin mRNA content increases to a maximal value in the fractions rich in basophilic normoblasts of 30,000–50,000 molecules/cell. However, the accuracy of these determinations is not as great as for the ratio determinations, and no significant deviations were seen except in the cells rich in pronormoblasts, which contained less globin mRNA than the later stages.

scholarly journals Stability of alpha and beta globin messenger RNA during induced differentiation of mouse erythroleukemia cells

Blood ◽  
1979 ◽  
Vol 54 (4) ◽  
pp. 933-939
Author(s):  
R Gambari ◽  
RA Rifkind ◽  
PA Marks
Keyword(s):  
Messenger Rna ◽  
Erythroid Differentiation ◽  
Beta Globin ◽  
Globin Mrna ◽  
Induced Differentiation ◽  
Hexamethylene Bisacetamide ◽  
Erythroleukemia Cells ◽  
Mrna Content ◽  
Murine Erythroleukemia ◽  
Murine Erythroleukemia Cells

Murine erythroleukemia cells (MELC) are induced to express erythroid differentiation when cultured with hexamethylene bisacetamide (HMBA). Newly synthesized alpha and beta globin mRNA are both relatively stable, half-life (t1/2) greater than 50 hr, early in the course of induced differentiation. In fully induced cells there is a decrease in stability of both newly synthesized alpha and beta globin mRNA. The decay of alpha mRNA is faster, (t 1/2, 10--12 hr) than beta globin mRNA (t1/2, 20--22 hr). Thus, differences in stability of alpha and beta globin mRNA plays a role in determining the ratio of alpha to beta mRNA content in differentiated erythroid cells.

scholarly journals A specific dimerization of rabbit β-globin messenger ribonucleic acid

1978 ◽  
Vol 175 (1) ◽  
pp. 159-169 ◽  
Author(s):  
V A Mezl ◽  
J A Hunt
Keyword(s):  
Beta Globin ◽  
Globin Mrna ◽  
Cross Hybridization ◽  
Rapid Preparation ◽  
Free System ◽  
Messenger Ribonucleic Acid ◽  
Low Salt ◽  
Alpha Globin ◽  
Two Peaks ◽  
Second Peak

Rabbit globin mRNA, when layered in low salt on 0.1 M-NaCl/sucrose gradients, separates into two peaks of material. Translation of these two RNA fractions in the wheat-germ cell-free system, hybridization against globin complementary DNA (cDNA) and cross-hybridization against cDNA species prepared from each fraction show that the first peak sedimenting at 10S is a alpha-globin mRNA and the second peak, sedimenting at approx. 15S, is beta-globin mRNA. The sedimentation rate of the beta-globin mRNA is concentration-dependent. By changing concentration and pH, it is indicated that in low-salt beta-globin mRNA adopts a conformation that leads to specific, but weak, self-dimerization during centrifugation in 0.1M-NaCl. This property permits rapid preparation of intact and relatively pure alpha- and beta-globin mRNA species.

scholarly journals Stability of alpha and beta globin messenger RNA during induced differentiation of mouse erythroleukemia cells

Blood ◽  
1979 ◽  
Vol 54 (4) ◽  
pp. 933-939 ◽  
Author(s):  
R Gambari ◽  
RA Rifkind ◽  
PA Marks
Keyword(s):  
Messenger Rna ◽  
Erythroid Differentiation ◽  
Beta Globin ◽  
Globin Mrna ◽  
Induced Differentiation ◽  
Hexamethylene Bisacetamide ◽  
Erythroleukemia Cells ◽  
Mrna Content ◽  
Murine Erythroleukemia ◽  
Murine Erythroleukemia Cells

Abstract Murine erythroleukemia cells (MELC) are induced to express erythroid differentiation when cultured with hexamethylene bisacetamide (HMBA). Newly synthesized alpha and beta globin mRNA are both relatively stable, half-life (t1/2) greater than 50 hr, early in the course of induced differentiation. In fully induced cells there is a decrease in stability of both newly synthesized alpha and beta globin mRNA. The decay of alpha mRNA is faster, (t 1/2, 10--12 hr) than beta globin mRNA (t1/2, 20--22 hr). Thus, differences in stability of alpha and beta globin mRNA plays a role in determining the ratio of alpha to beta mRNA content in differentiated erythroid cells.

The structure of pre-mRNA and mRNA from erythroid cells

1978 ◽  
Vol 36 (2) ◽  
pp. 234-235
Author(s):  
A.-M. Ladhoff ◽  
B.J. Thiele ◽  
Ch. Coutelle ◽  
S. Rosenthal
Keyword(s):  
Bone Marrow ◽  
Structural Transformation ◽  
Electron Micrographs ◽  
Ammonium Acetate ◽  
Bone Marrow Cells ◽  
Erythroid Cells ◽  
Globin Mrna ◽  
Ordered Structures ◽  
Rabbit Bone ◽  
Rabbit Reticulocytes

The suggested precursor-product relationship between the nuclear pre-mRNA and the cytoplasmic mRNA has created increased interest also in the structure of these RNA species. Previously we have been published electron micrographs of individual pre-mRNA molecules from erythroid cells. An intersting observation was the appearance of a contour, probably corresponding to higher ordered structures, on one end of 10 % of the pre-mRNA molecules from erythroid rabbit bone marrow cells (Fig. 1A). A virtual similar contour was observed in molecules of 9S globin mRNA from rabbit reticulocytes (Fig. 1B). A structural transformation in a linear contour occurs if the RNA is heated for 10 min to 90°C in the presence of 80 % formamide. This structural transformation is reversible when the denatured RNA is precipitated and redissolved in 0.2 M ammonium acetate.

scholarly journals Blood transcriptome analysis of patients with uncomplicated bacterial infection and sepsis

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Velma Herwanto ◽  
Benjamin Tang ◽  
Ya Wang ◽  
Maryam Shojaei ◽  
Marek Nalos ◽  
...  
Keyword(s):  
Bacterial Infection ◽  
Healthy Controls ◽  
Beta Globin ◽  
Transcriptome Data ◽  
Valuable Resource ◽  
Globin Mrna ◽  
Blood Samples ◽  
Data Description ◽  
Pathway Activation ◽  
Blood Transcriptome

Abstract Objectives Hospitalized patients who presented within the last 24 h with a bacterial infection were recruited. Participants were assigned into sepsis and uncomplicated infection groups. In addition, healthy volunteers were recruited as controls. RNA was prepared from whole blood, depleted from beta-globin mRNA and sequenced. This dataset represents a highly valuable resource to better understand the biology of sepsis and to identify biomarkers for severe sepsis in humans. Data description The data presented here consists of raw and processed transcriptome data obtained by next generation RNA sequencing from 105 peripheral blood samples from patients with uncomplicated infections, patients who developed sepsis, septic shock patients, and healthy controls. It is provided as raw sequenced reads and as normalized log2 transformed relative expression levels. This data will allow performing detailed analyses of gene expression changes between uncomplicated infections and sepsis patients, such as identification of differentially expressed genes, co-regulated modules as well as pathway activation studies.

Polyomavirus enhancer contains multiple redundant sequence elements that activate both DNA replication and gene expression

1985 ◽  
Vol 5 (4) ◽  
pp. 649-658
Author(s):  
G M Veldman ◽  
S Lupton ◽  
R Kamen
Keyword(s):  
Dna Replication ◽  
Globin Gene ◽  
Mrna Levels ◽  
Beta Globin ◽  
Transcriptional Enhancer ◽  
Globin Mrna ◽  
Enhancer Region ◽  
Sequence Elements ◽  
Multiple Copies ◽  
A Site

Sequences that comprise the 244-base-pair polyomavirus enhancer region are also required in cis for viral DNA replication (Tyndall et al., Nucleic Acids Res. 9:6231-6250, 1981). We have studied the relationship between the sequences that activate replication and those that enhance transcription in two ways. One approach, recently described by de Villiers et al. (Nature [London], 312:242-246, 1984), in which the polyomavirus enhancer region was replaced with other viral or cellular transcriptional enhancers suggested that an enhancer function is required for polyomavirus DNA replication. The other approach, described in this paper, was to analyze a series of deletion mutants that functionally dissect the enhancer region and enabled us to localize four sequence elements in this region that are involved in the activation of replication. These elements, which have little sequence homology, are functionally redundant. Element A (nucleotides 5108 through 5130) was synthesized as a 26-mer with XhoI sticky ends, and one or more copies were introduced into a plasmid containing the origin of replication, but lacking the enhancer region. Whereas one copy of the 26-mer activated replication only to 2 to 5% of the wild-type level, two copies inserted in either orientation completely restored replication. We found that multiple copies of the 26-mer were also active as a transcriptional enhancer by measuring the beta-globin mRNA levels expressed from a plasmid that contained either the polyomavirus enhancer or one or more copies of the 26-mer inserted in a site 3' to the beta-globin gene. We observed a correlation between the number of inserted 26-mers and the level of beta-globin RNA expression.

scholarly journals The identification of globin messenger ribonucleic acid in newt erythropoietic cells

1979 ◽  
Vol 183 (1) ◽  
pp. 105-114
Author(s):  
J A Grasso ◽  
G P Casale
Keyword(s):  
Ionic Strength ◽  
Mrna Translation ◽  
High Ionic Strength ◽  
Globin Mrna ◽  
Triturus Cristatus ◽  
Messenger Ribonucleic Acid ◽  
Reticulocyte Lysate ◽  
Globin Synthesis ◽  
26S Rrna ◽  
Mrna Precursor

Polyadenylated [poly(A)+]-RNA isolated from newt (Triturus cristatus) erythropoietic cells contained two main species sedimenting at 9S and 25S, and minor amounts of a 15-20S component. The 9S poly(A)+-RNA fraction induced synthesis of newt haemoglobin and globins in frog oocytes and in an mRNA-dependent rabbit reticulocyte lysate, confirming its identity as newt globin mRNA. Translation of 9S globin mRNA in reticulocyte lysate was concentration-dependent, the patterns of globin synthesis suggesting both preferential utilization and unequal amounts of the different globin mRNA subspecies. Globin mRNA activity was also evident in the 25S poly(A)+-RNA fraction whose localization in polyribosomes excluded its function as a nuclear globin mRNA precursor. Denaturation in formamide and estimation of its relative methyl content indicated that the 25S poly(A)+-RNA fraction contained equimolar amounts of 9S globin mRNA and 26S rRNA. Translation of the 25S fraction in reticulocyte lysate was less efficient than that of comparable amounts of 9S globin mRNA and induced a pattern of globin synthesis similar to that obtained with subsaturating amounts of 9S mRNA. The 25S mRNA-rRNA complex was considered to be a non-physiological aggregate generated by extraction of RNA in the presence of buffers of moderate to high ionic strength.

PSVII-27 Genetic analysis of disease resilience of nursery-to-finish pigs under a natural disease challenge model using reaction norms

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 235-235
Author(s):  
Jian Cheng ◽  
KyuSang Lim ◽  
Austin Putz ◽  
Anna Wolc ◽  
John Harding ◽  
...  
Keyword(s):  
Genetic Model ◽  
Average Daily Gain ◽  
Genetic Correlations ◽  
Reaction Norm ◽  
Cubic Spline ◽  
Clinical Disease ◽  
Random Regression ◽  
Disease Phenotypes ◽  
Linear Reaction ◽  
Better Than

Abstract Disease resilience is the ability of an animal to maintain performance across environments with different disease challenge loads (CL) and can be quantified using random regression reaction norm models that describe phenotype as a function of CL. Objectives of this study were to: 1) develop measures of CL using growth rate and clinical disease phenotypes under a natural disease challenge; 2) evaluate genetic variation in disease resilience. Data used were late nursery and finisher growth rates and clinical disease phenotypes, including medical treatment and mortality rates, and subjective health scores, collected on 50 batches of 60/75 crossbred (LRxY) barrows under a polymicrobial natural disease challenge. All pigs were genotyped using a 650K SNP panel. Different CL were derived from estimates of contemporary group effects and used as environmental covariates in reaction norm analyses of average daily gain (ADG) and treatment rate (TRT). The CL were compared based on model loglikelihoods and estimates of genetic variance, using both linear and cubic spline reaction norm models. Linear reaction norm models fitted the data significantly better than the standard genetic model and the cubic spline models fitted the data significantly better than the linear reaction norm model for most traits. CL based on early finisher ADG provided the best fit for nursery ADG, while CL based on clinical disease phenotypes was best for finisher ADG and TRT. With increasing CL, estimates of heritability for ADG initially decreased and then increased, while estimates of heritability for TRT generally increased with CL. Genetic correlations were low between ADG or TRT at high versus low CL but high for close CLs. Results can be used to select more resilient pigs across different CL levels, or high-performance animals at a given CL level, or a combination of these. Funded by Genome Canada, Genome Alberta, USDA-NIFA, and PigGenCanada.

Characterization of the major regulatory element upstream of the human alpha-globin gene cluster

1991 ◽  
Vol 11 (9) ◽  
pp. 4679-4689
Author(s):  
A P Jarman ◽  
W G Wood ◽  
J A Sharpe ◽  
G Gourdon ◽  
H Ayyub ◽  
...  
Keyword(s):  
Regulatory Element ◽  
Globin Gene ◽  
Major Elements ◽  
Beta Globin ◽  
Globin Mrna ◽  
Expression Studies ◽  
Primary Element ◽  
Alpha Globin ◽  
Binding Sequence

The major positive regulatory activity of the human alpha-globin gene complex has been localized to an element associated with a strong erythroid-specific DNase I hypersensitive site (HS -40) located 40 kb upstream of the zeta 2-globin mRNA cap site. Footprint and gel shift analyses of the element have demonstrated the presence of four binding sites for the nuclear factor GATA-1 and two sites corresponding to the AP-1 consensus binding sequence. This region resembles one of the major elements of the beta-globin locus control region in its constitution and characteristics; this together with evidence from expression studies suggests that HS -40 is a primary element controlling alpha-globin gene expression.

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