Mechanisms of cell differentiation: Affinity of a morphogenetic factor to DNA

Summary A morphogenetic factor which induces inTriturus gastrula ectoderm tissues which are derived from mesoderm and endoderm has been extracted from chicken and amphibian embryos. The factor which is protein in nature has been obtained from chicken embryos in a highly purified state.The biological activity of the chicken factor is partially inhibited when the factor is combined with chicken DNA or sonicated chicken DNA.When the 3H-labelled factor is combined with sonicated DNA and then centrifuged on a sucrose gradient the factor migrates in part with the DNA. This indicates that the factor is bound to DNA.The inferences from these results are discussed with regard to the possible mechanism of action of the factor and the molecular mechanism of differentiation.     

The effect of reversal of Na and K electrochemical potential gradients on the active transport of amino acids in Ehrlich ascites tumor cells

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1. The net uptake of α-aminoisobutyric acid (AIB) in Ehrlich ascites tumor cells has been studied under a variety of transmembrane concentration gradients of Na⁺, K⁺ and AIB itself.     

Temperature-sensitive initiation of DNA replication in a mutant of Escherichia coli K12

Summary A mutant of E. coli K12 appears to be temperature-sensitive in the process of initiation of DNA replication. After a temperature shift from 33 to 42°C, the amount of residual DNA synthesis (Fig. 1) and the number of residual cell divisions (Figs. 2,4) indicate that rounds of DNA replication in process are completed, but new rounds cannot be initiated. Following the alignment of chromosomal DNA by amino acid starvation at 33° C no residual DNA synthesis at 42°C takes place (Fig. 5). When the temperature is lowered to 33°C after a period of inhibition at 42°C, the following observations are made: 1. DNA replication resumes and proceeds synchroneously, (Figs. 7, 8a), 2. cells start to divide again only after a lag period of about 1 hour 3. a temporary increase in cell volume is correlated with the frequency of initiation of DNA synthesis (Fig. 8a, b). In a lysogenic mutant strain prophage is inducible; with all bacteriophages tested, replication of phage DNA is not inhibited at 42°C.     

The fate of newly made DNA in Escherichia coli mutants thermosensitive in DNA synthesis

Summary E. coli mutants exist in which DNA synthesis is thermosensitive. In one class of these mutants DNA synthesis stops immediately if a critical temperature (42°C) is reached. When DNA replication in such mutants is followed by ³H thymidine incorporation at 33°C, it is found that 1. only the newly made DNA is degraded at 42°C, 2. the discontinuously replicated DNA is lost predominantly at 42°C, 3. 1–3% of the chromosomal DNA is rendered acid soluble at 42°C without concomitant loss of viability of the cells at 33°C.Replication of phage DNA is inhibited in the same mutant at 42°C. However, when DNA synthesis is followed in infected cells at 33°C it is found that 1. no degradation of specific DNA seems to occur at 42°C in the early phase of infection, 2. replicating DNA molecules in the late phase of infection are completed at 42°C before DNA synthesis comes to a halt.     

Ultrastructural localization of cell wall teichoic acids in Streptococcus faecalis by means of concanavalin A

Some teichoic acids are known to be partially substituted by α-D-glucopyranosyl residues such as the teichoic acids of Streptococcus faecalis NCIB 8191. They will, therefore, bind specifically the phytohemagglutinin concanavalin A. Concanavalin A labelled with mercury or colloidal gold coated with concanavalin A has been used to mark isolated cell walls in order to localize the teichoic acids at the ultrastructural level. Besides these two direct marking techniques, the indirect concanavalin A-peroxidase technique (localization of peroxidase by the diaminobenzidine method followed by postosmication) has been applied to thin sections of premarked cells. All three methods gave almost identical results, namely, a dense and homogeneous distribution of the cell wall teichoic acids. In control experiments total inhibition was achieved in the presence of methyl-α-D-mannopyranoside. After trichloroacetic acid or alkali extraction of the teichoic acids from isolated walls no marking could be detected.     

Aspekte der Siebröhren-Differenzierung bei Monocotylen

Zusammenfassung Ausgehend vom nachmeristematischen Zustand einer prospektiven Siebröhre machen Kern, Tonoplast, ER, Dictyosomen und Ribosomen wÄhrend der Zelldifferenzierung folgende Umwandlungen durch:DerKern erfÄhrt in seiner Matrix eine zunehmende Substanzverarmung, an deren Ende seine vollstÄndige Auflösung steht. In der Kernmembran sind lokale Erweiterungen des intrazisternalen Raumes charakteristisch für den Beginn der degenerativen VerÄnderungen. Stets bleibt die Kernhülle jedoch in lockerem Kontakt mit dem ER; vermutlich geht sie zum Abschlu\ der Kernauflösung in das Membransystem des ER ein.In AbhÄngigkeit von der Rückbildung des Zellsaftraums zerfÄllt derTonoplast zunÄchst in zisternenartige Abschnitte, um spÄter in der ausdifferenzierten Siebröhre ganz zu verschwinden. Wie an verschiedenen Beispielen abzulesen ist, werden diese VerÄnderungen offensichtlich auf dem Höhepunkt der protoplasmatischen Differenzierung eingeleitet.Das gleichmÄ\ig granulÄr-zisternaleEndomembransystem junger Siebelemente — hÄufig charakterisiert durch spiralig aufgereihte Ribosomen — wandelt sich mit zunehmender Hydratisierung des Protoplasten in tubulÄre bis vesikulÄre Elemente um und geht zum anderen über verschiedene Zwischenstufen in gitterartige Membranstrukturen ein.Die in der jungen Siebröhre zahlreichen, polar gebautenDictyosomen tragen mit der Produktion von Golgi-Vesikeln zum Wandaufbau bei. Im Laufe der Zelldifferenzierung werden in steigendem Ma\e auch coated vesicles abgegeben, deren Bedeutung noch unklar ist. In der ausdifferenzierten Siebröhre sind weder Dictyosomen noch die von ihnen abgegebenen Vesikel nachzuweisen. Dieses Schicksal teilen sie mit denRibosomen, die gleichfalls nur in jungen Siebelementen anzutreffen sind und dort zu Polysomen vereint, hÄufig in spiraliger oder helixförmiger Anordnung vorkommen.

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Aspects of sieve-tube differentiation in monocotyledons

Summary Subsequent to the postmeristematic state of a future sieve-tube its nucleus, tonoplast, endoplasmic reticulum, dictyosomes, and ribosomes undergo the following transformations:Thenucleus shows a gradually declining ground substance until at last it entirely disintegrates. Local enlargements of the intracisternal space of the nuclear membrane indicate the beginning of the degenerative phase. During all these alterations as well as in the meristematic state the nuclear envelope is in loose contact with the endoplasmic reticulum. We suppose that it finally becomes part of a complex ER system. Subject to the disorganization of the central cell-sap room thetonoplast initially removing from the parietal protoplast later on completely disappears. According to our results these transformations are obviously introduced at the climax of protoplasmatic differentiations.The uniform rough surfaced cisternal ER of young sieve elements—often characterized by spirally arranged ribosomes—changes during sieve-tube ontogeny to tubular or vesicular elements and partly fuses to lattice-like membrane structures passing several other stages of membrane condensation.Polardictyosomes that are numerous in young sieve-tubes produce coated vesicles besides smooth golgi-vesicles.Ribosomes are abundant in young elements, too, they are often found as polysomes of helical or spiral arrangement. Neither dictyosomes nor ribosomes have been seen in differentiated sieve-tubes.

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Teil einer Habilitationsschrift der Math.-Naturw. FakultÄt Bonn.