Effects of the mutant gene wellhaarig in chimeric mice

The mutant gene wellhaaring (we) confers the waved coat in mice, which is most pronounced in homozygotes at 10 to 21 days of postnatal development. Abnormal hair growth and structure in the we/we mutant mice results from defective cell differentiation in the inner root sheath of a hair follicle. To localize the site of the we gene action, we obtained ten chimeric mice by aggregation of the early C57BL/6-2we/we and BALB/c embryos. The chimera coat was waved, shaggy, or almost normal depending on the percentage of the mutant component. In the we/we +/+ chimeric animals of the first generation (G1) aged 21 days, both mutant and normal hair phenotypes were observed, which was especially discernible in zigzag hair. Note that none of the chimeras exhibited the alternating patterns of transversely oriented stripes or patches of either mutant or normal hair; i.e., they had a mixed parental hair phenotype. We also did not observe the animals with an intermediate phenotype, which suggests a discontinuous hair formation in chimeras according to the "all or nothing" principle. The data obtained indicate that the dermal papilla cells of a hair follicle are the sites for the we gene action. During the embryonic development, dermal cells are strongly mixed, which accounts for the lack of the clear-cut transverse stripes of either mutant or normal hair. The mutant gene we is probably responsible for a disrupted induction signal from the dermal papilla towards ectodermal cells of a hair follicle.     

The Effects of Mutant Gene hairless in Chimeric Mice

The autosomal recessive gene hairless (hr) is responsible for the complete hairlessness in mice homozygous for this gene. Hair shedding that begins at the age of 10 days is caused by an abnormal cycle of hair follicle development disturbed at the catagen stage. This results in enhanced programmed cell death (apoptosis) and ultimately leads to the complete hair follicle destruction and shedding of all hairs by the age of three weeks. To study the phenotypic expression of the hr gene in a chimeric organism, we have obtained 12 chimeric mice hr/hr +/+ by means of aggregation of early embryos hr/hr and +/+. In chimeric mice, the hair shedding has begun two days later than in the hr/hr mice. By day 23 of postnatal development, hairless areas were present on the coat of chimeric mice or the latter were completely hairless depending on the percentage of the hr/hr mutant component. In four chimeras with high content of the mutant component (68–76%), the hair shedding process was similar to that in the hr/hr mice, though it was accomplished two days later. In three chimeras with 48–51% of the mutant component, alternating hairless and hair-covered bands were observed. These data suggest that the hr gene acts in epidermal cells of a hair follicle, because epidermal cell clones in embryonic skin migrate in the lateral–ventral direction coherently and without mixing. However, some chimeras displayed a pattern which was not so clear-cut: the band borders were illegible and hairs partly covered the hairless areas. In some chimeras, the uniform thinning of the coat was observed. Analysis of the effects of the hr mutant gene in chimeric mice differing in the ratio between mutant (hr/hr) and normal (+/+) components in tissues suggests that the hrgene acts in the epidermal cells of the hair follicle. The interactions between cells have an essential effect on the mode and degree of the hr gene expression, which leads to distortion of the ectodermal coat pattern in chimeras.     

Morphogenesis of the hair follicle during catagen

Summary During catagen, the transition period between growth and quiescence, the growing (anagen) hair follicle is reorganized to form the resting (telogen) follicle. The last portion of the hair shaft produced at the onset of catagen consists only of cortex. Surrounding the cortex and attached tightly to it are the club cells, which resemble the cortex in structure and development except that the filaments of the club are oriented randomly and do not exhibit the keratin pattern seen in the cortex. The club cells in turn are attached to a capsule of germ cells which are formed by progressive transformation of the outer root sheath cells at the middle of the growing follicle. When the capsule of germ cells is formed, the follicle below it undergoes resorption, presumbaly mediated by hydrolytic enzymes. As the follicle disintegrates, the surrounding basal lamina undergoes extensive pleating and is eventually resorbed. Collagen fibers around the basal lamina are engulfed and degraded by the large number of macrophages that surround the hair follicle at this time. The dermal papilla remains as a compact ball of cells just below the capsule of germ cells.This study constitutes publication No. 428 from the Oregon Regional Primate Research Center, supported by Grant No. FR-00163.I wish to thank Mrs. Janice Anderson for patient and excellent technical assistance and Mr. Joel Ito for the drawing.     

Outer root sheath (ORS) cells organize into epidermoid cyst-like spheroids when cultured inside Matrigel: a light-microscopic and immunohistological comparison between human ORS cells and interfollicular keratinocytes

In organotypic cultures, outer root sheath (ORS) cells of the human hair follicle develop into a stratified epithelium largely reminiscent of the epidermis; this apparently reflects their importance during wound healing. In the present study, ORS cells were grown inside a three-dimensional network of extracellular matrix proteins (Matrigel), together with different mesenchymal cells, in an attempt to mimic their follicular environment. Thus, inside Matrigel, ORS cells formed spheroids differentiating toward the center and showing all the markers of epidermal keratinization. Under identical conditions, normal epidermal keratinocytes developed similar spheroids, but of a significantly smaller size. Human dermal fibroblasts and dermal papilla cells, cocultured in the matrix, had a positive influence on both the proliferation and differentiation within both types of spheroids. Epidermal differentiation markers, such as suprabasal keratins, involucrin, filaggrin, gp80 and pemphigoid antigen, were readily expressed in ORS spheroids, whereas hard (hair) keratins were not detectable by immunostaining. Cells positive for an epithelial membrane antigen, strongly expressed in sebaceous glands, were seen in numerous spheroids. In contrast to organotypic surface epithelia, the expression and location of different integrin chains was normalized in ORS spheroids, indicating an enhanced mesenchymal influence in this in vitro system.     

Effects of mutations in genesfadR,fabB, fadE,andenvC ofEscherichia coli on the action of the lysis gene of bacteriophageϕX174

The lytic effect of the expression of the cloned geneE of bacteriophage X174 inEscherichia coli is considerably amplified by a mutation in thefadR gene, which primarily affects the regulation of fatty acid degradation. In contrast, reduction of the fluidity of the cell membranes by use of thefabB andfadE mutations, which interfere with the synthesis and the oxidation of unsaturated fatty acids, severely inhibits the action of the X174 lysis gene product. A chain-forming mutant carrying a pleiotropic mutation in theenvC locus is also refractory to the X174 lysis protein. As shown by reversion and complementation of theenvC mutatation, a defect in at least one additional gene (rle) is involved in the generation of this refractoriness.     

Biochemische Untersuchungen zur Frage der Existenz eines “silent Gene” im Polymorphismus der Pseudocholinesterasen

Zusammenfassung Zwei Seren des Phänotypus Ch₁SS, die unter Standardtestbedingungen keine Pseudocholinesteraseaktivität aufwiesen, wurden mit Hilfe der Stärkegel-Elektrophorese, der Mikromanometrie und mit immunologischen Methoden näher untersucht.Nach elektrophoretischer Auftrennung läßt sich in diesen silent gene-Seren eine Pseudocholinesteraseaktivität nachweisen; im Vergleich zu der Aktivität von Normalserum erscheint diese sehr gering und läßt sich nur in der c₄-Zone erkennen. Die Identifizierung gelingt mit Hilfe spezifischer Färbeverfahren unter Verwendung verschiedener Substrate und Inhibitoren.Die quantitative Bestimmung von Pseudocholinesteraseaktivität im silent gene-Serum mit mikromanometrischen Methoden ergab eine Aktivität von 2–3% gegenüber den Kontrollen (Benzoylcholin als Substrat).Durch Immunisierung von Kaninchen mit gereinigtem Pseudocholinesteraseprotein wurden Antiseren erhalten; zwischen silent gene-Serum und diesen Antiseren konnten Präzipitationsreaktionen im Immuno-Diffusionstest, in der Immuno-Elektrophorese und mit einer Immuno-Adsorptionsmethode nachgewiesen werden. Diese Ergebnisse lassen annehmen, daß bei den von uns untersuchten Fällen das silent gene im Pseudocholinesterasepolymorphismus eine Enzymproteinsynthese steuert; das nachgewiesene Pseudocholinesteraseprotein scheint sich qualitativ von dem Enzymprotein des Normalserums zu unterscheiden.

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Two sera without pseudocholinesterase activity corresponding to the homozygous phenotype Ch₁SS are examined by electrophoretical, manometric, and immunological methods.These silent gene sera show no activity under the common conditions (spectrophotometric assay).After electrophoretical separation of silent gene serum an esterase activity is found which can be identified as pseudocholinesterase activity, although it is weak in comparison with the activity of usual sera. The pseudocholinesterase activity of silent gene serum can be demonstrated only in the zone c₄ where 90% of the total activity is present if usual serum is inserted. The identification has been achieved by staining procedures applying several substrates and inhibitors.Quantitative estimation of this pseudocholinesterase activity was carried out by micromanometric assays with benzoylcholine as substrate. The activity of silent gene sera was 2–3% of normal serum.Antisera against human pseudocholinesterase-protein have been obtained by immunization of rabbits with a highly purified enzyme protein. Between these antisera and the homozygous silent gene sera precipitates were found in immuno double-diffusion tests and immunoelectrophoresis. They could be identified as pseudocholinesterase protein by esterase staining under various conditions.Quantitative estimations have been carried out by immuno-adsorption assays comparing the amount of antibody fixed by usual serum and by silent gene serum.The results presented in this paper suggest that the silent gene in pseudocholinesterase polymorphism induces in these two cases the synthesis of an enzyme protein which is similar to the enzyme protein of usual pseudocholinesterase. The weak activity is due to a qualitative difference between silent gene enzyme protein and the normal pseudocholinesterase protein. A structural alteration of the enzyme protein is assumed to be more likely than a quantitative difference in protein synthesis.

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Einige der Ergebnisse dieser Arbeit wurden auf dem 2nd Meeting of the Federation of European Biochemical Societies, Wien 1965, vorgetragen.

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Wesentliche Teile dieser Arbeit werden von R. A. Hofmann der Medizinischen Fakultät der Universität Freiburg i. Br. als Inauguraldissertation vorgelegt.     

The we Gene is a Modifier of the walGene in Mice

Interaction of gene wellhaarig (we) with genes waved alopecia(wal) and hairless (hr) was studied in mice. The mutant gene weis responsible for the development of a specific waved coat in homozygotes. Homozygous mice carrying mutant gene walalso have a wavy coat, though a partial alopecia develops with time in these animals. In homozygotes for thehr gene, hair loss is observed beginning from the age of ten days. A series of crosses we/weand wal/wal yielded animals with we/+wal/wal and we/we wal/wal genotypes. In micewe/+wal/wal carrying gene we at a single dose, alopecia is accelerated significantly as compared to the single-dose homozygotes +/+wal/wal. In we/we wal/wal mice, alopecia starts earlier than in we/+wal/wal mice; by the age of one month, the double homozygotes are almost hairless except for small body areas covered with a sparse coat. In addition, curliness of the first-generation hair in mice we/we wal/wal is much more expressed than in +/+wal/wal and we/we+/+ mice. The obtained evidence suggests that the wegene is a modifier of the wal gene because the former enhances the effects of the walgene, which is confirmed by the earlier onset of alopecia and progression of the latter in mice having the we/+wal/wal genotype and especially in we/we wal/wal animals. The we/we hr/+ mice do not differ in coat from we/we+/+ mice; in both cases, the coat is wavy. The coat of double homozygotes we/we hr/hr, is similar to that of we/we+/+ mice until ten days of age, when the signs of alopecia appear. By the age of 21 days, mice we/we hr/hr have lost their coat completely like mice +/+ hr/hr. Hence, the we gene is a modifier of the walgene though it does not interact with hrgene during the coat formation.     

Transgenic markers for mammalian chimeras

Summary We describe the construction of aggregation chimeras between normal and transgenic embryos containing multiple copies of mouse -globin genes. The transgenic component of the chimeras is then detected in tissue sections by a DNA-DNA in situ hybridization technique, using a biotinylated DNA -globin probe and an avidin-linked alkaline phosphatase detection system. The general advantages of transgenic markers for chimeras are discussed.     

A maternal-effect mutation disturbs extracellular matrix organization in the early Pleurodeles waltl embryo

Summary In the early development of the urodele amphibian Pleurodeles waltl, a fibronectin-containing extracellular matrix underlies the inner face of the blastocoel roof. When gastrulation occurs, the fibronectin fibrils provide a suitable substrate for mesodermal-cell migration. Delay in morphogenetic movements of gastrulation has been described in embryos from mutant females (ac/ac) of Pleurodeles waltl. Studies of abnormal mutant gastrulae with fluorescent lectins and immunostaining for fibronectin reveal that they lack a normal matrix. The fibronectin-containing extracellular material always gives rise to a granular pattern without fibronectin-fibril formation. Fibronectin and ₅₁ syntheses occur normally in maternal-effect embryos. In vitro, mesodermal cells from early mutant gastrulae adhere and migrate on fibronectin-conditioned substrata.     

The<Emphasis Type="Italic"> ssgB</Emphasis> gene,encoding a member of the regulon of stress-response sigma factor σ<Superscript>H</Superscript>, is essential for aerial mycelium septation in<Emphasis Type="Italic"> Streptomyces coelicolor</Emphasis> A3(2)

The Streptomyces coelicolor A3(2) gene ssgB belongs to the regulon of stress-response sigma factor ^H. By integrative transformation via double cross-over, a stable null mutant of ssgB was obtained. This mutation had no obvious effect on vegetative growth, but critically affected aerial mycelium septation. The S. coelicolor ssgB mutant produced aerial hyphae without any signs of septation into spore compartments. The mutation was complemented in trans by wild-type ssgB including the ^H-dependent ssgBp promoter. The results proved that ssgB belongs a developmental branch of the ^H regulon.     

Isolation and characterization of an ω3-desaturation-defective mutant of an arachidonic acid-producing fungus,Mortierella alpina 1S-4

A mutant considered to be defective in the conversion of n-6 to n-3 fatty acids (3-desaturation) was derived from a 5-desaturation-defective mutant (Mut44) of Mortierella alpina 1S-4, after treating its spores with N-methyl-N-nitro-N-nitrosoguanidine. This mutant cannot produce 8(Z),11(Z),14(Z),17(Z)-eicosatetraenoic acid or any other n-3 fatty acids, of which about 10% was found in its parental strain upon cultivation at 12°C. The mutant's growth rate was comparable to that of the parental strain when grown at 28°C, but it became much slower when the mutant grew at 12°C, at which the lag phase for Mut44 was about 2 d but 5 d for the mutant.Abbreviations 18:33 9(Z),12(Z),15(Z)-octadecatrienoic acid - 18:43 6(Z),9(Z),12(Z),15(Z)-octadecatetraenoic acid - 20:43 8(Z),11(Z),14(Z),17(Z)-eicosatetraenoic acid - AA arachidonic acid - DHGA dihomo--linolenic acid - EPA 5(Z),8(Z),11(Z),14(Z),17(Z)-eicosapentaenoic acid - GLC gas-liquid chromatography - MNNG N-methyl-N-nitro-N-nitrosoguanidine - PC phosphatidylcholine     

Cytoplasmic male sterility in barley

Summary The effect of the msm1 cytoplasm of barley (Hordeum vulgare L.) on kernel protein and lysine was studied using the near-isogenic, unrestored derivatives of seven barley varieties. With normal lysine varieties, Adorra, Bomi, CI 4362, and Hankkija's Eero, the msm1 cytoplasm produced an average of one percentage point more protein than did the normal cytoplasm of the same varieties. There was no difference between the two cytoplasms with respect to their effect on the lysine content. With high lysine varieties, Bomi Risø mutant 13, Bomi Risø mutant 1508, and CI 3947, msm1 produced almost one percentage point more protein but protein with a somewhat decreased lysine content.Induced partial spike fertility in normal Adorra was found to be associated with lysine in meal (r=–0.999), with protein in meal (r=–0.984), and with lysine in protein (r=0.941). Removal of the spikes on the secondary tillers affected both the protein and its lysine content. It is suggested that good spike fertility is an important pre-requisite when selecting high lysine and/or high protein segregants or mutants.     

Eucampia Index as an indicator of the Late Pleistocene oscillations of the winter sea-ice extent at the ODP Leg 119 Site 745B at the Kerguelen Plateau

A new paleoenvironmental proxy, Eucampia Index, was used to trace the Late Pleistocene oscillations of winter ice extent at ODP Leg 119, Site 745B (59° 35.71 S and 85° 51.60 E) on the Kerguelen Plateau. The index is calculated as the ratio of winter terminal to intercalary valves of the diatom Eucampia antarctica sensu lato. During the early Brunhes the winter sea-ice edge was positioned south from Site 745. It started expanding northward, closer to the site location soon after 0.4 Ma and progressed in a manner of a several wide oscillations. For approximately the last 0.1 Ma the winter sea-ice edge oscillated less and retained similar range of oscillations. The ice edge oscillated in periods which correspond closely to that of Milankovitch oscillations of Earth obliquity, although the significance of individual periods appears to vary in time.     

Novel seed lipid phenotypes in combinations of mutants altered in fatty acid biosynthesis inArabidopsis

Summary The seed fatty acid (FA) composition of various single mutant combinations ofArabidopsis thaliana that affect FA biosynthesis has been examined. Double mutant combinations offae, a mutation affecting CIS elongation, and a series of four other FA biosynthetic mutants were synthesized. The four other single mutants were:fad2 andfad3, which are deficient in 181 and 182 desaturation, respectively;fab1, which is elevated in 160 and decreased in 181; andfab2, which is elevated in 180 and decreased in 181. The superimposition of two blocks in the FA biosynthetic pathway leads to dramatic changes in the FA content of the double mutants. The tenArabidopsis stocks analyzed to date (wild-type, five single mutants, and four double mutants) make seed oils with a wide range of FA compositions, and illustrate the diversity of oils it is possible to obtain from a single plant species.     

Proriatic hair follicle cells

Calmodulin levels were measured by radioimmuno-assay in freshly isolated and cultured psoriatic human scalp hair follicle cells. The mean value ± SEM for calmodulin was 1.97±0.15 ng calmodulin g^-1 protein for 16 control subjects whereas calmodulin levels were significantly increased in psoriatic hair follicles, 2.93±0.26 ng calmodulin g^-1 protein (uninvolved skin) for 18 patients and 3.09±0.21 ng calmodulin g^-1 protein for involved skin derived hair follicles for 17 of these patients. In vitro, 3-week-old cultures of psoriatic keratinocytes contained less DNA and more calmodulin per DNA than their normal counterparts. When 6 week-old cultures of psoriatic and control hair follicle keratinocytes were compared, this difference disappeared. These results are related to the state of differentiation of these cultures.     

Lambda phage mutants insensitive to temperature-sensitive repressor

Summary Weak-virulent mutants of temperate coli-phage were isolated which can grow on the CIts lysogen producing a temperature-sensitive repressor but which cannot grow on the wild type lysogen producing a normal repressor.Genetic analysis on the mutants shows that their weak-virulence is attributable to two mutations, one (virL) in the region between sus N₂₁₃ and c ₄₇ and the other (virR) in the region between c ₁ and sus O₈. Both mutations are located within the region of non-homology between and imm ⁴³⁴ phages.True virulent mutants which can grow on the wild type lysogen can be obtained easily from the weak-virulent mutant by an additional mutation, virC in a region very close to virR. The virulent mutants obtained are similar to the classical vir mutant (Jacob and Wollman, 1954). The virL and virR mutations are probably operator mutations which render the genome insensitive to the repressor.This work was reported at the XII th International Congress of Genetics, held in Tokyo, on August 23, 1968.     

Phyllotaxis,anthotaxis and semataxis

Summary Long-lasting debates, caused by conflicting viewpoints among biometrists on the phenomena of rhythmic growth in plant shoots, are at last being settled on certain crucial points. Most workers today agree that not all symmetrical constructions in plants can be explained by the application of the phyllotaxis theory. This theory explains adequately the orthostichous arrangement of leaves on growing photosynthetic apices, but fails in the case of non-photosynthetic reproductive organs.In the present paper three successive systems of shoot arrangement are described: phyllotaxis for cauline leaves, anthotaxis for sporophylls, and semataxis for semaphylls (see Table II). Each of these systems serves a different purpose: photosynthesis, reproduction and advertisement, and is accordingly adapted to its special function. Phyllotaxis and anthotaxis are classical concepts dealing with the arrangement of leaves and flowers; semataxis is a new term used in this paper to describe the arrangements of semaphylls. Goethe's classical theory of the metamorphosis of plants (1790) and his hypothetical Urpflanze are discussed in this paper from the viewpoint of the phyllotaxis-semataxis relation.

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Zusammenfassung Nach den langen Debatten über die Anwendbarkeit der Phyllotaxistheorie zur Erklärung der Blatt- und Blütenstellungen, hat man sich schliesslich über einige alte Streitfragen geeinigt. Man nimmt nun ganz allgemein an, dass nicht alle symmetrische Bildungen an Pflanzen sich mit der Anwendung der Phyllotaxisgesetze und Fibonazzi Reihe begründen lassen. Diese Theorie erklärt glaubwürdig die rhythmische Anordnung der vegetativen Blätter an den photosynthetischen Sprossen, nicht aber die symmetrischen Bildungen in den Blüten.In dem vorliegenden Aufsatz sind drei historisch nacheinander folgenden Entwicklungssysteme: Phyllotaxis, für die vegetativen Blätter, Anthotaxis für Sporophyllen und Semataxis für die Semaphyllen eingehend diskutiert worden. Phyllotaxis ist ein klassischer Begriff für die Anordnung der Blätter; Anthotaxis wurde später eingeführt, ist bis jetzt aber nur wenig in diesem Sinne gebraucht und deshalb in diesem Aufsatz neu definiert worden. Semataxis ist neu. Sie betrifft die Anordnung der Semaphyllen, der auffälligen Teile (meistens die farbigen Kronblätter oder die kronblätterartige Kelch- und Hochblätter) der von den Insekten bestäubten Blüten der höheren Pflanzen. Jedes von diesen Systemen dient einem bestimmten Zweck. Die phyllotaktische Anordnung der vegetativen Blätter bezweckt die rationellste Photosynthese. Anthotaxis ist die zweckmässigste Ordnung der Sporophyllen für die mechanische Bestäubung und Fruchtbildung, während Semataxis schliesslich die Ankündigung der insektenbestäubten Blüten bedeutet. Alle diese Systeme sind ihren besonderen Funktionen eingehend angepasst.Obwohl die historische Entwicklung der Sprossysteme der höheren Pflanzen nach der Reihenfolge: Phyllotaxis Anthotaxis Semataxis erfolgt, sind in dem vorliegenden Aufsatz mehrere Ausnahmen mit umgekehrter Richtung angeführt. Semataktische Bautypen können unmittelbar von Phyllotaxis, und anthotaktische Anordnungen von Semataxis abgeleitet werden.Ferner sind in diesem Aufsatz die klassischen Theorien vonGoethe über die Metamorphose der Pflanzen (1790) und die hypothetische Urpflanze von dem neuen Gesichtswinkel der Phyllotaxis- und Semataxislehre eingehend diskutiert worden.

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Sommaire Après de longs débats sur le phénomène de la croissance rhythmique des pousses des plantes, les biométristes s'accordent aujourd'hui sur certains points essentiels de la discussion. Au contraire des auteurs antérieurs, les chercheurs d'aujourd'hui admettent que toutes les constructions symétriques des plantes ne s'expliquent pas par l'application de la théorie de phyllotaxie. Cette théorie explique suffisamment la disposition orthostichique des feuilles sur des sommets photosynthétiques croissants, mais échoue dans le cas des organes reproductifs non-photosynthétiques.Dans le présent écrit trois systèmes successifs d'arrangements de pousses sont décrits: phyllotaxie pour feuilles caulines, anthotaxie pour sporophylles et sémataxie pour sémaphylles (voir table II). Chacun de ces systèmes sert à des fins différents: à la photosynthèse, à la reproduction et à l'avertissement, et s'adapte par conséquent à sa propre fonction.Phyllotaxie et anthotaxie sont des conceptions classiques pour désigner la disposition des feuilles et des fleurs, sémataxie est un term nouveau qui sert à désigner dans cette étude l'arrangement des sémaphylles.La théorie classique deGoethe sur la métamorphose des plantes (1790) et son hypothétique Urpflanze sont discutées dans cet écrit du point de vue de la relation existant entre la phyllotaxie et la sémataxie.

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The author is indebted to Dr. W. H.Bragonier, Head of the Department of Botany and Plant Pathology for financial support of this project.

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Journal Paper No. J — 3561 of the Iowa Agricultural and Home Economics Experiment Station, Ames, Iowa. Project No. 1073.     

Variations in the rate of synthesis of beta and beta'' RNA polymerase polypeptides under the influence of certain factors.

Summary In merodiploid cells containing a double dose of structural genes of RNA polymerase subunits-rpoBand rpoC-the rate of and subunits synthesis is 2 times higher than in haploidcells. Missence mutation rpoC1 (tsX) alters polypeptide and inducesthe and subunits synthesis at increased rate, particularly at a nonpermissive temperature. When rpoBCoperon carrying mutation rpoC1 is duplicated no dosage effect is observed. In the rpoC ⁺/rpoC1 heterodiploid the rpoC1 mutation does not significantly accelerate RNA polymerase subunits synthesis i.e. is recessive with respect to rpoC ⁺ Rifampicin causes 6-fold stimulation of RNA polymerase subunits synthesis in a sensitive wild-type strain. The rpoC1 mutation itself accelerates the synthesis of these subunits 3-fold. In the presence of rifampicin the mutant strain produces 13–22-fold faster as compared to wild-type strain without the drug. Thus, the effects of rifampicin and the mutation are multiplied suggesting that these factors act independently. Similar data have also been obtained with rifampicin-treated cells of rpoB22 (ts22) amber-mutant.After UV-irradiation of cells and synthesis is depressed much stronger than the total protein synthesis. Infection with a transducing phage rif ^d-47 which carries rpoB gene provokes a higher rate of synthesis. When pre-irradiated cells (500 erg/mm²) are infected with this phage, the rate of synthesis grows 20-fold compared to irradiated, non-infected cells and 6.5-fold compared to intact cells.The data are discussed in terms of the possible regulatory mechanisms of RNA polymerase subunit synthesis.     

LH-producing cells in the ovine pituitary

Summary An immunocytochemical technique using the peroxidase-antiperoxidase complex (PAP) was applied to identify and characterize the LH-secreting cells in the ovine pituitary at the ultrastructural level. These cells, round or oval in shape, possessing flattened cisternae of the rough endoplasmic reticulum, contain one class of secretory granules (mean diameter 250 nm) and large dense bodies (600 to 800 nm in diameter). LH molecules and the two subunits LH and LH were localized on the secretory granules and on the small granules near the Golgi complex. The large dense bodies, the cisternae of the endoplasmic reticulum and the saccules of the Golgi complex showed no reaction product.Abbreviations used in this Article O-LH ovine luteinizing hormone - b-LH bovine luteinizing hormone - p-LH porcine luteinizing hormone - p-LH porcine LH subunit - p-LH porcine LH subunit - O-FSH ovine follicle stimulating hormone - b-TSH bovine thyrotropic hormone - A-b LH antiserum to bovine LH - A-pLH antiserum to porcine LH subunit - A-pLH antiserum to porcine LH subunit     

Control of sporulation in yeast: SAD1-a mating-type specific, unstable alteration that uncouples sporulation from mating-type control

Summary Normally, in order to sporulate, a diploid yeast cell must be heterozygous (MAT a/MAT) at the mating-type locus. In a new mutant, this requirement is circumvented by SAD1. This alteration is mating-type specific; it allows sporulation of MAT/ MAT. MAT/mat a-1, and MAT/mat-2 diploid cells, but not MAT a/MAT a or MAT a/0 (monosomic) strains. Other than acquiring the ability to sporulate, SAD1 cells behave as wild-type MAT/MAT strains; they exhibit medial budding, normal mating factor production and response and mate with normal mating efficiencies and kinetics. The segregation of SAD1 is often bizarre; for example, MAT/MAT strains which were constructed to be heterozygous SAD1/+ often segregate 4 SAD1:0+progeny and strains which were constructed to be homozygous SAD1/ SAD1 sometimes segregated 1 SAD1:3+progeny. The genetic analyses of SAD1 suggest that it is dominant and is located 30 cM from MAT on chromosome III.