Phosphorylation of Chromosomal Proteins in Resting and Wounded Potato Tuber Tissues

Wounding of quiescent white potato tuber tissue enhances chromatin-boundprotein phosphokinase activity, which exhibits two distinctphases during wound-healing. A moderate activation of the enzymesup to 20 hr after injury is followed by a dramatic increasein activity with a peak at 50 hr. This time-course resemblesthat of chromatinbound DNA-dependent RNA polymerase with a peakin activity at about 48 hr after wounding. The kinases phosphorylateendogenous proteins as well as added histones, phosvitin andcasein. The incorporated phosphate is stable under standardassay conditions, indicating the absence of protein phosphatases.Sensitivity of the incorporated phosphate toward trypsin andalkali, but not DNase, RNase, hydroxylamine or succinic acidpoints to seryl- and threonyl-bonds and proteins as acceptormolecules. Kinases from resting tissues are only weakly stimulatedeven by 100 mM MgCl₂, those from wounded tissues exhibit pronouncedMg^$$-optima at 5–10 mM with endogenous proteins, phosvitinand casein and 50 mM MgCl₂ with histones. Wounding also increasesthe sensitivity of the kinases toward p-hydroxymercuribenzoate. Chromatin preparations from both resting and wounded tissuescontain about 40 protein bands after polyacrylamide disc gelelectrophoresis. In vitro phosphorylation of these proteinsin chromatin from quiescent tissues is comparably low and uniform.Wounding induces changes in the protein and phosphorylationpattern with a general enhancement of phosphorylative capacityand preferential phosphorylation of low molecular weight proteins. (Received August 10, 1981; Accepted November 18, 1981)     

Decreased fertility in related females heterozygous for the 1/29 chromosome translocation

Eighteen heifers and 120 cows which were descendants of a presumed 1/29 carrier Simmental bull were karotyped. Nine heifers (50%) and 48 cows (40%) were found to be heterozygous for the 1/29 translocation (59, XX, t(1q;29q)). The other animals were chromosomally normal (i.e., 60, XX) or not karotyped. The 48 1/29 cows were compared with 72 chromosomally normal cows with regards to days to first conception, calving interval, percentage of calves conceived, percentage of calves weaned and production efficiency (% calved conceived × % calved weaned). Nine carrier heifers were compared to the nine noncarrier heifers as to pregnancy status. Carrier, noncarrier and nonkarotyped relatives were compared to each other and to contemporary females with regard to pregnancy status at their initial exposure to males. The percentage of calves conceived (calving efficiency) in the 72 noncarrier and the 48 females heterozygous for the 1/29 translocation were 81.5 and 74.8%, respectively (P<0.07). Although days to first conception was longer and percentage of calves weaned and production efficiency were lower in the female heterozygous for the 1/29 translocation, the differences were not statistically different (P>0.10) from the noncarriers. Pregnancy rate was 44.4 and 66.7% (P>0.10) for nine carrier and nine noncarrier heifers, respectively. The pregnancy rate of carrier (65.4%), noncarrier (73.2%) and nonkarotyped (77.8%) relatives of this sire at their mating as yearlings, did not differ (P>0.10). The pregnancy rate as yearlings of carrier females (65.4%) and contemporary heifers (79.8%) did differ (P<0.05). Comparing the pregnancy rate as yearlings of all descendants (72.0%) of the Simmental sire to contemporary heifers (79.8%), a significant decrease (P<0.05) was found indicating that fertility of this sire may have been lower than other sires or that other factors beside the translocation affected fertility.     

Diversity of the mouse T cell receptor Cγ1 gene: structural analysis in C57BL/Ka

We have isolated an unusual T cell receptor chain cDNA clone (7.1) from a library made from RNA derived from adult thymus of C57BL/Ka mice. This cDNA clone corresponds to the appropriately processed C1 constant region exons preceded by 1.5 kb of J-C1 intron. The 7.1 coding region is extremely homologous to the C1 gene of BALB/c mice, differing at the protein level by a single deletion (alanine 139) and a single substitution. This latter change eliminates the sole N-linked sugar attachment site, providing a basis for strain-specific glycosylation patterns. The J-C1 intronic region contains two DNA segments (termed J1 and J2) that are highly reminiscent of joining (J) segments; both have potentially functional recombination and donor splice sequences flanking an open reading frame. Northern analysis suggests that 7.1 may be derived from a large, variable region-containing precursor.     

Interactions of porphyrins with purified DNA and more highly organized structures

Studies of the solution properties of gold(III)tetrakis(4-N-methylpyridyl) porphine and its DNA binding characteristics have been conducted utilizing uv/vis absorption spectroscopy, circular dichroism (CD), Mossbauer spectroscopy, and temperature-jump relaxation techniques. These studies indicate that over the concentration range considered this water soluble gold(III) porphyrin does not aggregate, binds axial ligands only weakly with a preference for soft Lewis bases, and is capable of intercalation into nucleic acids of appropriate base pair content. The interaction of this and several other porphyrins with the synthetic polynucleotide poly(dA-dC).poly(dT-dG) has been studied. Spectroscopic signatures for intercalation were found for those derivatives not having axial ligands. Intercalation into chromatin in vitro can also occur with those porphyrins and metalloporphyrins which do not have axial ligands. Finally, studies utilizing microinjection techniques indicate that once within the cell, tetrakis(4-N-methylpyridyl)porphine tends to localize in the nucleus.     

Differential regulation and tissue-specific distribution of enzymes of phenylpropanoid pathways in developing parsley seedlings

Characteristic enzymes of general phenylpropanoid metabolism (phenylalanine ammonialyase) and of the flavonoid-glycoside and furanocoumarin branch pathways (chalcone synthase and S-adenosyl-l-methionine: bergaptol O-methyltransferase, respectively) were localized immuno-histochemically in cross-sections of various aerial parts of parsley (Petroselinum crispum) at different stages of seedling development. Phenylalanine ammonia-lyase occurred predominantly in epidermal and oil-duct epithelial cells, but was also detectable in other tissue parts. The two pathway-specific enzymes were localized in the epidermis (chalcone synthase) and in oil ducts (bergaptol O-methyl-transferase). High chalcone-synthase concentrations occurred very early in leaf development and then declined. High levels of the methyltransferase were present at all times investigated. The temporal and spatial at all times investigated. The temporal and spatial distribution of all three enzymes is in agreement with the time courses and sites of accumulation of the biosynthetic end products.Abbreviations BMT S-adenosyl-l-methionine: bergaptol O-methyltransferase - CHS chalcone synthase - PAL phenylalanine ammonia-lyase     

Divergent changes in the concentrations of gonadotropin beta-subunit messenger ribonucleic acid during the estrous cycle of sheep

Cyclic changes in the production of the pituitary gonadotrophic hormones, LH and FSH are essential events in the maintenance of the reproductive system of female mammals. While studies have examined changes in the secretion of LH and FSH during the estrous cycle and demonstrated the importance of these hormones in regulation of ovarian development and gametogenesis, considerably less is known concerning the regulation of the biosynthesis of these hormones. Although initial studies have examined changes in LH subunit mRNA concentrations during the rat and ovine estrous cycles, no information concerning the physiological regulation of FSH beta mRNA concentrations has been available. In the present study we have examined the relationship between pituitary concentrations of LH and FSH subunit mRNAs and the serum concentrations of these gonadotropins. The results demonstrate a very different pattern of change for FSH beta subunit mRNA than that observed for alpha and LH beta subunit mRNAs. In fact, FSH beta mRNA concentration decline substantially during the preovulatory period, reaching minimal values at a time when alpha and LH beta mRNA levels are near maximal. Furthermore, this decline in FSH beta mRNA amounts occurs when serum FSH concentrations are maximal. Thus, FSH beta mRNA concentrations follow a very different pattern than that of serum FSH. In contrast, LH beta mRNA and serum LH concentrations tend to increase at the same time. These findings provide evidence that concentrations of LH beta and FSH beta mRNAs are likely regulated by different mechanisms.