Cis-trans test shows a functional relationship between non-allelic lethal mutations in the T/t-complex

Recessive lethal mutations in the T/t-complex of the mouse characteristically show defective genetic complementation, even when they affect very different stages of embryogenesis and are known to be nonallelic. To address the question of their genetic or functional relationship, we have applied the cis-trans test, using several well defined recombinant t-chromosomes that carry two or more lethal mutations, and others that are devoid of specific lethals. We show here that the defective complementation that occurs between different t-lethals is a specific result of the trans configuration; thus these genes, which may map as much as 15 cM apart, constitute a functional unit. Some speculations are presented to interpret this enigma in terms of DNA plasticity.     

Structural correlates of cellular tumorigenicity and anchorage independence in transformed fibroblasts

The ability on nonhematopoietic cells to proliferate in vitro without attachment to a solid surface (anchorage independence) is known to be highly correlated with their ability to form tumors in nude mice. Transformed cells in vitro frequently also show less organization of the intracellular actin-containing microfilament bundles and less cell-surface fibronectin compared to normal cells. We have examined whether the loss of the anchorage requirement for growth is related to either of these structural changes in the cellular cytoskeleton. Our approach was to select a series of subclones from a nontransformed Syrian hamster fibroblast line, NIL8, for the acquisition of either anchorage independence in vitro or tumorigenicity in nude mice. These subclones were then examined for concomitant changes in the cytoskeletal structures. We found that anchorage independence, decreased actin cable organization, and tumorigenicity in nude mice were coordinately induced in both the in vitro- and in vivo-selected subclones, whereas the loss of fibronectin was not consistently coinduced with these three markers. These results suggest that the transformation-associated decrease in actin cable organization in this type of cell enhances the ability to grow without anchorage in vitro and to form tumors in vivo.     

Direct control by calcium of 25-hydroxycholecalciferol-1-hydroxylase activity in chick kidney mitochondria

Kidney mitochondria were isolated from rachitic chicks and their activity in the metabolism of 25-OH-D₃ was studied in relation to the amount of calcium added $\text{in vitro}$. The addition of 0.050.2 mM calcium to a mitochondrial suspension caused a marked and dose-related stimulation of 1-hydroxylation. A sharp decline in the activity was induced by higher concentrations (0.3-0.5 mM) of calcium. The rate of 24-hydroxylation was not influenced by calcium. In these effects, calcium was relatively specific among various divalent cations. These data strongly suggest that calcium is directly involved in the regulation of the vitamin D activation in kidney mitochondria.     

Interaction of metal ions with polynucleotides and related compounds. XXII. Effect of divalent metal ions on the conformational changes of polyribonucleotides

Changes in the conformation of poly(G), poly(C), poly(U), and poly(I) in the presence of divalent metal ions Mg²⁺, Ca²⁺, Mn²⁺, Co²⁺, Ni²⁺, Cu²⁺, Cd²⁺, and Zn²⁺ have been measured by means of ORD and u.v. spectra. Mg²⁺ and Ca²⁺ ions stabilize helical structures of all the polynucleotides very effectively at concentrations several orders of magnitude lower than the effective concentration of Na⁺ion. Cu²⁺ and Cd²⁺ destabilize the helical structure of polynucleotides to form random coils. Zn²⁺, Ni²⁺, Co²⁺, and Mn²⁺ions do not behave in such a clear-cut manner: they selectively stabilize some ordered structures, while destabilizing others, depending on the ligand strength of the nucleotide base as well as the preferred conformation of that polynucleotide.