Identification of ‘Amigo’ and ‘Kavkaz’ translocations in Ohio soft red winter wheats (Triticum aestivum L.)

One cultivar (GR876) and two advanced Ohio soft red winter wheat lines (OH413 and OH414), with Kavkaz in their pedigrees, were examined for the presence of the Kavkaz, 1RS/1BL rye/wheat chromosome translocation. Another advanced line (OH416), with Amigo in its pedigree, was examined for the presence of the Amigo, 1RS/1AL translocation. Only two satellited chromosomes were observed in most mitotic root-tip cells from GR876, OH413, and OH414, compared to four in most cells from OH416. Heteromorphic bivalents were observed in most PMCs from hybrids produced by crossing GR876, OH413, and OH414 as females to Chinese Spring. No heteromorphic bivalents were observed in PMCs from OH416 x Chinese Spring hybrids. When GR876 and the Ohio lines were hybridized with Chinese Spring dimonotelosomic-1B, telosomic trivalents, consisting of the short- and longarm telosomes paired with chromosome 1B, were only observed in PMCs from 43-chromosome hybrids involving OH416. The long-arm telosome paired with the translocation chromosome, while the short-arm telosome remained unpaired in all other 43-chromosome hybrids. Separation of gliadin proteins from GR876 and the Ohio lines by PAGE revealed that secalin bands for GR876, OH413, and OH414, migrated similarly to the secalins for Kavkaz. Bands for OH416, identified as possible secalins, migrated similarly to those for Amigo. Cultivar GR876 and advanced Ohio soft red winter wheat lines OH413 and OH414 carry the Kavkaz translocation, while OH416 carries the Amigo translocation.Communicated by K. Tsunewaki     

Characterization and transcript analysis of the major phycobiliprotein subunit genes from Aglaothamnion neglectum (Rhodophyta)

The genes encoding the and subunits of allophycocyanin, phycocyanin and phycoerythrin from the red alga Aglaothamnion neglectum were isolated and characterized. While the operons containing the different phycobiliprotein genes are dispersed on the plastid genome, the genes encoding the and subunits for each phycobiliprotein are contiguous. The subunit gene is 5 for both the phycocyanin and phycoerythrin operons, while the subunit gene is 5 for the allophycocyanin operon. The amino acid sequences of A. neglectum phycobiliproteins, as deduced from the nucleotide sequences of the genes, are 65–85% identical to analogous proteins from other red algae and cyanobacteria. The conserved nature of the plastid-encoded red algal and cyanobacterial phycobiliprotein genes supports the proposed origin of red algal plastids from cyanobacterial endosymbionts.Many environmental factors effect phycobilisome biosynthesis. The effect of both nutrient availability and light quantity on the level of A. neglectum phycobiliprotein subunits and the mRNA species encoding those subunits is described.     

“Differentiation Induction” culture of human leukemic myeloid cells stimulates high production of macrophage differentiation inducing factor

A suitable procedure for the production of human monokines was defined as differentiation-induction culture. Human monocytic leukemia THP-1 cells were well-differentiated from nonfunctional promonocytes into macrophage-like cells by the induction with a combination of mezerein, retinoic acid, and aMycoplasma fermentans extract. The differentiated THP-1 cells secreted a high amount of macrophage differentiation-inducing factor (DIF) activity and concomitantly produced other known monokines, such as tumor necrosis factor- (TNF-), interleukin-1 (IL-1) and interleukin-1 (IL-1), into the medium. These results suggest that other novel human monokines may also be found in the conditioned medium of THP-1 cells induced by the differentiation-induction culture conditions defined in this study. Macrophage DIF was purified to homogeneity and NH₂-terminal amino acid sequence analysis revealed that macrophage DIF is very similar or identical to human leukemia inhibitory factor (LIF). The cDNA encoding human LIF was isolated using the polymerase chain reaction, and a clone producing 3.7 g/10⁶ cells day recombinant LIF was selected from Chinese hamster ovary (CHO) cells which were transfected with the LIF cDNA. The recombinant LIF production in CHO cells was quantified using MTT reduction assay with M1 cells.     

\4 integrin expression onin vitro andin vivo metastatic variants of lewis lung carcinoma

The expression of 4, 6, and 1 integrin subunits has been investigated on somein vitro andin vivo murine metastatic variants derived from Lewis lung carcinoma (3LL). By the use of monoclonal antibodies which recognizes different epitopes of 6, 1, and 4 subunits we demonstrate that 6 and 1 subunits are expressed in all metastatic variants of 3LL irrespective of their metastatic potential, whereas 4 subunit is expressed only in highly metastasizing cells of 3LL. Northern blots of different metastatic variants probed with 1 and 4 subunits demonstrate thata) significant amounts of 1 mRNA were detected in all metastatic variants of 3LL;b) mRNA corresponding to the described entire coding sequence of 4 subunit is expressed only on highly metastasizing cells of 3LL. We conclude that 4 subunit is specifically expressed in highly metastasizig cells of 3LL while is undetectable in lower metastasizing ones.     

Association of Polymorphic Marker A1/A2 of Gene ITGB3 with Coronary Artery Disease and Myocardial Infarction

The distributions of the alleles and genotypes of polymorphic marker A1/A2 of gene ITGB3 encoding the 3 subunit of integrin IIb3 in groups of patients with coronary artery disease (CAD), including those who have had myocardial infarction (MI), and in the control group have been compared. Analysis using Fisher's test has not revealed significant differences between these groups with respect to the distributions of the alleles and genotypes of polymorphic marker A1/A2 of gene ITGB3. This indicates that this polymorphic marker is not associated with either CAD or MI in the Moscow population.     

Intermolecular contact sites in protein kinase CK2

Chemical crosslinking and analysis of CNBr-digested fusion products by immunoblotting with sequence-specific antibodies identifies an interaction between positions 55-70 of subunit (55-70) and 65-80 of subunit (65-80). This has been supported by crosslinking of subunits with peptides 65-80 and 55-70, by binding of subunits to immobilized peptides, and by the hindrance of coprecipitation with peptide-raised antibodies (anti-65-80; anti-55-70). Functionally, 55-70 is a negative regulatory region for the kinase activity of subunit . The opposite, stimulatory property of subunit has been assigned to its C-terminal part. Subdivision of peptide 155-181, that has stimulatory effect, into overlapping peptides and assaying for a binding and binding competition revealed a tight physical contact at 162-175. This region, however, is non-stimulatory indicating binding a necessary but not sufficient quality for stimulation. A contact might exist to regions surrounding C147 and/or C220 at subunit a as indicated by crosslinking and peptide competition. The crosslinking data also confirm a - contact in CK2 holoenzyme. Effects by non-ionic detergents show hydrophobic interactions to play an important role in catalytic activity adjustment.     

Modification of domains of α and β subunits of F1-ATPase from the thermophylic bacterium PS3, in their isolated and associated forms,by 3′-O-(4-benzoyl)benzoyl adenosine 5′-triphosphate (BzATP)

Photoaffinity labeling by 3-O-(4-benzoyl)benzoyl adenosine 5-triphosphate (BzATP) of the adenine nucleotide binding site(s) on isolated and complexed and subunits of F₁-ATPase from the thermophilic bacterium PS3 (TF₁) is described. BzATP binds to both isolated and subunits, to complexed subunit but not to complexed subunit. Amino acid sequence determination of radiolabeled peptides obtained by proteolytic digestion of [-³²P]BzATP-labeled subunit indicates that residues on both the amino-terminal (residues A41-E67) and carboxy-terminal (residues Q422–Q476) were modified by BzATP. One of the residues in the carboxy-terminal modified by BzATP is most probably Q422. Although the binding stoichiometry of 1 mol of BzATP incorporated by either isolated or complexed subunit was maintained, the spatial conformation of the polypeptide determines which amino acid residue(s) is more accessible to the reactive radical. CNBr derived fragments G10-M64, E75-M233, and D390-M469 were labeled with the isolated subunit. With complexed subunit the label was found only in CNBr fragments: E75-M233 and G339-M389. The locations where the covalently bound BzATP was found, in the soluble and assembled subunits, indicate that different conformational states exist. In the isolated form of the and subunits the amino- and carboxy-termini can fold and reach the central domain of the polypeptide, the domain containing the adenine nucleotide binding site. When combines with to form the ₃₃ core complex the new conformation of the subunits is such that covalent labeling by BzATP of and of the amino terminal of subunit is excluded.     

Effects of triiodothyronine (T3) on differentiation of rat cortical neurons in primary cultures

Some of the events which characterize neuronal terminal differentiation have been studied in rat cortical neurons cultured in a selective synthetic medium for a period which corresponds to terminal brain maturation in vivo. In particular, we have studied the effect of T3 on the synthesis of nuclear proteins and the expression of the mRNAs which encode different variants of T3 nuclear receptors (c erb A proteins). We have shown that: a) T3 stimulates the turnover of nuclear proteins, with a more evident effect on the non-histone component; b) for the whole lifespan of cultures the predominant form of c erb A mRNA is the 2 variant (which encodes a protein unable to bind T3); whatever the function of 2 protein this finding suggests that its predominance on 1 is settled very early during mammalian brain maturation.     

Interaction sites on phosphorylase kinase for calmodulin

Holophosphorylase kinase was digested with Glu-C specific protease; from the peptide mixture calmodulin binding peptides were isolated by affinity chromatography and identified by N-terminal sequence analysis. Two peptides originating from the subunit, having a high tendency to form a positively charged amphiphilic helix and containing tryptophane, were synthesized. Additionally, a homologous region of the subunit and a peptide from the subunit present in a region deleted in the isoform were also selected for synthesis. Binding stoichiometry and affinity were determined by following the enhancement in tryptophane fluorescence occurring upon 1:1 complex formation between these peptides and calmodulin. Finally, Ca²⁺ binding to calmodulin in presence of peptides was measured. By this way, the peptides 542–566, 547–571, 660–677 and 597–614 have been found to bind specifically to calmodulin.Together with previously predicted and synthesized calmodulin binding peptides four calmodulin binding regions have been characterized on each the and subunits. It can be concluded that endogenous calmodulin can bind to two calmodulin binding regions in as well as to two regions in and . Exogenous calmodulin can bind to two regions in and in . A binding stoichiometry of 0.8mol of calmodulin/ protomer of phosphorylase kinase has been determined by inhibiting the ubiquitination of calmodulin with phosphorylase kinase. Phosphorylase kinase is half maximally activated by 23nM calmodulin which is in the affinity range of calmodulin binding peptides from to calmodulin. Therefore, binding of exogenous calmodulin to activates the enzyme. A model for switching endogenous calmodulin between , and and modulation of ATP binding to as well as Mg²⁺/ADP binding to by calmodulin is presented.