Histone acetylation and heterochromatin content of cultured Peromyscus cells

In order to explore the relationship between unacetylated arginine-rich histones and condensed chromatin structure, the extent of histone acetylation was examined in cultured cell lines derived from three species of deer mice. These species differ considerably in their genomic content of heterochromatin but contain essentially the same euchromatin content. Cells of Peromyscus eremicus, containing 34–36% more constitutive heterochromatin than Peromyscus boylii or Peromyscus crinitus cells were found to contain 28–35% more unacetylated histone H4, 22–29% more unacetylated histone H3, and 18–22% more unacetylated histone H2B. This relationship between unacetylated histones and heterochromatin content was further explored by inducing hyperacetylation of P. eremicus and P. boylii histones through treatment of cells with 15 mM sodium butyrate for 24 h. It was found that the percentages of unacetylated histones H3 and H4 remaining after butyrate treatment were proportional to the amount of constitutive heterochromatin in the genome. These data support the concept that a small core of histones in constitutive heterochromatin is inaccessible to acetylation. It was also found that the acetylated state of isolated histones was sensitive to the method of histone extraction. Thus concern must be given to preparative procedures when studying histone acetylation in order to minimize these acetate losses.     

A surprising new protein superfamily containing ovalbumin,antithrombin-III,and alpha1-proteinase inhibitor

A distant relationship between chicken ovalbumin and two human plasma protease inhibitors was revealed by computer analyses. We propose a new protein superfamily containing at least three families: ovalbumin (and probably gene X and gene Y proteins), antithrombin-III, and alpha₁-proteinase inhibitor. Although these families may have diverged from a common ancestor more than 500 million years ago, they may still share similarity in gene structure as well as in protein sequence.     

The effects of antimicrotubule agents on cell motility in fibroblast aggregates

Spherical aggregates of chick heart, sclera and skin fibroblasts were fused with tritiated thymidine-labelled aggregates of the identical cell type. After being placed in contact, the two aggregates cohered and broadened the area of contact to form a single aggregate with a planar interface between the labelled and unlabelled halves. The motility of cells in the aggregate was determined by measuring the movement of labelled cells across the boundary into the unlabelled half. Exposure to pharmacological doses of antimicrotubule agents resulted in a significant reduction in fibroblast motility in the three-dimensional aggregates.     

Bromoacetylcholine as an affinity label of the acetylcholine receptor from Torpedo californica

Bromoacetyl[methyl-³H]choline is a highly specific label for the reduced acetylcholine binding site on the acetylcholine receptor from $\text{Torpedo californica}$. Only one of two binding sites per receptor monomer is susceptible to labeling. The labeled site is on the α chain of the receptor.     

Development of wheat bulb fly (Della coarctata fall.) larvae and pupae at different temperatures

When eggs of wheat bulb fly were added to young winter wheat plants in pots and kept at 5°, 8°, 10°, 12°, 15° and 20°, only 14%–44% of the eggs added produced larvae that succeeded in invading plants, but when newly hatched larvae were added directly 54%–92% were successful. At temperatures from 5° to 25°, the number of days required for larvae to complete their development in well-grown plants ranged from 11–14 days at 25° to 55–80 days at 5°. This corresponds to an accumulated temperature of approximately 286 day degrees C above the developmental threshold of 0.5°.The daily rate of development of larvae was marginally most rapid between 12° and 18°. When larvae were feeding in small, poorly growing plants, development was delayed and more shoots were needed before pupation than for healthy plants.The pupal stage which develops in the field from the end of April to June needs approximately 400 and 420 day degrees for completion in males and females (threshold 5°). Total day degrees required for the hatching of the egg to adult are about 700 (males) and 720 (females).Records of maximum and minimum temperatures from a meteorological screen 1 m above the ground can be used to calculate the number of day degrees accumulated from 1 May to 12 June, from which the dates of peak emergence of wheat bulb fly can be predicted. If 350 day degrees or more accumulate during this period, the peak emergence occurs near 20 June and with 200 or fewer day degrees it occurs near 11 July.The amount of damage to wheat by wheat bulb fly larvae in 1953, 1954, 1965 and 1966 depended largely on the number of eggs laid, the date of sowing, and also on the rate at which temperature accumulated in the autumn and winter; in all years, late sown crops would have had little opportunity to grow beyond the susceptible stage by the time they were attacked.

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Résumé Quand des jeunes plants de blé d'hiver en pots sont contaminés avec des oeufs de Delia coarctata et maintenus à des températures respectives de 5°, 8°, 10°, 12°, 15° et 20° C, seulement 14% à 44% de ces oeufs donnent des larves qui parviennent á attaquer les plants, alors que si on utilise directement des larves nouveau-nées on obtient 54 à 92% d'attaques réussies. Aux températures de 5° à 25° le temps nécessaire aux larves pour achever leur développement dans les plants bien développés avant de gagner le sol pour la pupaison, est de 11 à 14 jours à 25° contre 55 à 80 jours à 5°.Une courbe de la vitesse de développement établie à partir de la durée du stade larvaire de l'éclosion à la pupaison aux differentes températures montre que le développement est le plus rapide entre 12° et 18° et que le zéro de développement est aux alentours de 0,5°. Le nombre de degrésjours accumulés par la larve de l'éclosion à la pupaison est d'environ 286. Quand les larves se nourrissent aux dépens de plants petits à croissance faible, leur développement est retardé et exige plus de temps et plus de plantes avant la pupaison que lorsqu'elles se nourrissent sur des plants vigoureux.Le stade pupal qui intervient dans la nature de fin avril à juin, nécessite approximativement 400 à 420 degrés-jours pour le développement du mâle et de la femelle au-dessus d'une supposée température de base de 5°. Le nombre total de degrés-jours de l'oeuf à l'adulte est respectivement de 686 et 706 pour le mâle et la femelle.Les enregistrements des températures maxima et minima dans un abri météorologique à 1 m audessus du sol peuvent être utilisés pour établir le nombre de degrés-jours accumulés du 1 er'mai au 12 juin; à partir de ces données les pics d'émergence de D. coarctata peuvent être prédits. Si 350 degrésjours ou plus sont accumulés durant cette période, le pic d'émergence apparaît vers le 20 juin, mais si 200 degres-jours au moins sont notés le pic sera proche du 11 juillet.L'importance des dégâts occasionnés au bleî d'hiver en Angleterre par les larves de D. coarctata en 1953, 1954, 1965 et 1966 dépend largement du nombre d'oeufs pondus, de la date du semis et de la température accumulée en automne et en hiver; au cours de chacune de ces années les cultures à semis tardif auraient eu peu de possibilité de dépasser le stade sensible avant le moment où elles ont été attaquées.

     

Induction by ouabain of hemoglobin synthesis in cultured friend erythroleukemic cells

Induction of erythroid differentiation in ouabain-resistant murine erythroleukemia cells by ouabain is reported. Ouabain induction results in the appearance of hemoglobin-containing cells 12–24 hr earlier than induction of the same clone by dimethyl sulfoxide. The levels of globin mRNA after ouabain induction are similar in amount to the globin mRNA levels observed after induction by dimethyl sulfoxide. The concentration of ouabain required to induce hemoglobin synthesis depends upon the K⁺ ion levels in the culture medium. Lowering the extracellular K⁺ ion concentration 2–4 fold reduced by 10–40 fold the ouabain concentration necessary for the induction of hemoglobin synthesis. In low K⁺ medium (1.8 mM), ouabain is an effective inducer of hemoglobin synthesis at a concentration of 0.02 mM. This K⁺ effect is specific for ouabain induction, since induction by other inducers, such as dimethyl sulfoxide and dimethyl acetamide, does not exhibit this marked sensitivity to the levels of K⁺ ions in the culture medium. These results suggest that the binding of ouabain to the plasma membrane enzyme, $\text{Na}\text{K}$ ATPase, is required for the induction of erythroid differentiation by ouabain. A small but significant proportion of wild-type, ouabain-sensitive cells also can be induced by ouabain, below ouabain concentrations that are toxic to these cells. The observation that the binding of ouabain to the $\text{Na}\text{K}$ ATPase induces hemoglobin synthesis suggests that changes in the intracellular concentration of K⁺ ions may be involved in the control of erythroid differentiation in Friend erythroleukemic cells.     

Ovary-autonomous maternal inheritance at a developmental locus in Drosophila

The sex-linked temperature-sensitive mutation shibire^ts of Drosophila melanogaster shows a maternal effect causing embryonic lethality at 29°C. The maternal influence is due to gene action autonomous to the ovary. Embryos carrying the paternally derived wild-type gene can survive at 29°C but only if heat pulses are begun at least 9 hr after oviposition. The paternal rescue is presumably due to zygotic gene action at this locus beginning part way through embryogenesis. A maternal wild-type genome, however, can produce shi embryos that have sufficient shi⁺ product to support embryogenesis up to the hatching stage even at 29°C.     

Effect of L-azetidine 2-carboxylic acid on growth and proline metabolism in Escherichia coli

The effects of L-azetidine 2-carboxylic acid on growth and proline metabolism in a proline-requiring auxotroph of Escherichia coli are described. The homologue inhibited growth of the wild type and it, alone, did not substitute effectively for proline as a growth supplement for the mutant. In medium containing 0.05 mM proline, the addition of increasing amounts of homologue progressively inhibited growth of the wild type but stimulated growth of the mutant at homologue: proline ratios of 10 : 1 and 50 : 1. This suggested that the homologue exerted a “sparing effect” on proline in the mutant.The incorporation of L-[U-¹⁴C]proline and L-[³H]azetidine 2-carboxylic acid into hot trichloroacetic acid-insoluble material in the mutant was measured. Amino acid analysis of the insoluble material from cells incubated with radiolabeled proline alone revealed that proline was partially degraded and metabolized to other amino acids prior to incorporation into protein. The addition of unlabeled homologue to the incubation medium significantly reduced proline catabolism, suggesting that the homologue exerted a sparing effect on proline in this mutant. In medium containing unlabeled proline and radiolabeled L-azetidine 2-carboxylic acid, the homologuewas incorporated both intact and partially degraded prior to incorporation into protein. Alanine was the major L-azetidine 2-carboxylic acid catabolite.