Mechanism of desensitization of adenylate cyclase in lutropin. GTP-dependent uncoupling of the receptor

Lutropin-sensitive adenylate cyclase ((EC 4.6.1.1) ATP pyrophosphate-lyase (cyclizing)) in purified rat ovarian plasma membranes is stimulated by lutropin 2- to 3-fold in the absence, but 15- to 20-fold in the presence of GTP or p(NH)ppG. Following 10 to 15 min of incubation at 30 degrees C in the presence of lutropin, enzyme activity declined (50%) in the presence of GTP but not in the presence of p(NH)ppG. This desensitizing process induced by lutropin and GTP is not seen if NaF is also included in the incubation medium. The desensitized state of the enzyme persists at 4 degrees C in membranes washed free of the incubation medium. In this state the enzyme is characterized by: (i) a reduced response to lutropin even in the presence of p(NH)ppG; (ii) its response to NaF is not different from that of untreated enzyme; (iii) it reconverts to a fully responsive state following incubation (10 min, 30 degrees C) in GTP-free medium, a process accelerated by p(NH)ppG; (iv) the receptor content as well as the stability of the receptor.hormone complex does not differ from that of untreated fully responsive enzyme. It is proposed that desensitization results from a GTP-dependent, hormone-stimulated reaction that leads to impaired coupling of the enzyme system. The desensitized state induced is transient and may revert to a responsive one under specified conditions.     

Bacilliform inclusions in cells of the proximal pars distalis in the pituitary of four species of Tilapia (Teleostei)

Summary Thin sections and replicas of freeze-etched pituitaries from six species of the teleostean family of Cichlidae were studied by electron microscopy. Four species belonging to the genus Tilapia exhibit rod-like structures, i.e., bacilliform inclusions (BI), about 1–2 m in length in cells of the proximal Pars distalis. The BI are found either isolated or fused in small groups. They are enclosed in an envelope similar to that of secretory granules. Both the BI and the secretory granules give a positive PAS-reaction.Abbreviations BI bacilliform inclusion - GTH gonadotroph hormone - STH somatotroph hormone - TSH thyrotroph hormone This research was supported by a grant from the National Council for Research and Development, Israel, and the GKSS Geesthacht-Tesperhude, Germany     

The Routine Fitting of Kinetic Data to Models: A Mathematical Formalism for Digital Computers

A mathematical formalism is presented for use with digital computers to permit the routine fitting of data to physical and mathematical models. Given a set of data, the mathematical equations describing a model, initial conditions for an experiment, and initial estimates for the values of model parameters, the computer program automatically proceeds to obtain a least squares fit of the data by an iterative adjustment of the values of the parameters. When the experimental measures are linear combinations of functions, the linear coefficients for a least squares fit may also be calculated. The values of both the parameters of the model and the coefficients for the sum of functions may be unknown independent variables, unknown dependent variables, or known constants. In the case of dependence, only linear dependencies are provided for in routine use. The computer program includes a number of subroutines, each one of which performs a special task. This permits flexibility in choosing various types of solutions and procedures. One subroutine, for example, handles linear differential equations, another, special non-linear functions, etc. The use of analytic or numerical solutions of equations is possible.     

Determinants of site-specific recombination in the lambdoid coliphage HK022. An evolutionary change in specificity

The temperate bacteriophage HK022, like its relative lambda, inserts its chromosome into a specific site in the bacterial chromosome during lysogenization and excises it after induction. However, we find that the recombinational specificities of the two phages differ: they use different bacterial sites, and neither promotes efficient insertion or excision of the other phage chromosome. In order to determine the basis for this difference in specificity, we sequenced the HK022 elements that are involved in insertion and excision, and compared them to the corresponding lambda elements. The location, orientation, size and overall arrangement of the int and xis genes and the phage attachment sites are nearly identical in the two genomes, as is common for other functionally related elements in lambdoid phages. The Xis proteins of the two phages are functionally interchangeable, and their predicted amino acid sequences differ by but one residue. In contrast, the two Int proteins are not functionally interchangeable, and their sequences, although similar, differ at many positions. These sequence differences are not uniformly distributed: the amino-terminal 55 residues are completely conserved, but the remaining 302 show a pattern of differences interspersed with identities and conservative changes. These findings imply that the specificity difference between HK022 and lambda site-specific recombination is a consequence of the inability of the respective Int proteins to recognize pairs of heterologous attachment sites. The two phage attachment sites are remarkably similar, especially the two "arm" segments, which in lambda contain binding sites for Int, Xis and integration host factor. They are less similar in the segment between the two arms, which in lambda contains the points of recombinational strand exchange and a second class of binding site for Int protein (the "core-type" sites). The two bacterial attachment sites are quite different, although both have a short stretch of perfect homology with their respective phage partners at the points of strand exchange. We propose that the two Int proteins recognize similar or identical sites in the arms of their cognate attachment sites, and that differences in binding or action at the core-type sites is responsible for the divergent specificities. Genetic experiments and sequence comparisons suggest that both proteins recognize different but overlapping families of core-type sites, and that divergence in specificity has been achieved by an alternating succession of small, mutually compatible changes in protein and site.     

The frequency of two-base tracts in eukaryotic genomes

The frequency of two-base tracts is surveyed in a wide range of eukaryotic genomes using the special program TRACTS. All three two-base families are surveyed: R.Y (A,G.C,T), K.M (A,C.G,T), and S;W (A.T and G.C). Data for the human β-globin complex, for the tobacco chloroplast, and for 247 nt mammalian promoter regions are presented. All two-base tracts longer than three or four bases are overrepresented to an extent surpassing by far their occurrence in a randomized DNA population in the majority of the genomic regions analyzed; 20–30 long tracts are quite frequent, against the statistical odds. R.Y tracts are found at the largest excess, K.M tract to a slightly lesser extent, while S.W tracts are found at a moderate yet significant excess. The majority of the tracts manifest only a limited extent of tandem repeat structures. The idea that the two base tracts serve as unwinding elements is considered. Preseented at the NATO Advanced Research Workshop onGenome Organization and Evolution, Spetsai, Greece, 16–22 September 1992     

Changes in rat milk quantity and quality due to variations in litter size and high ambient temperature.

The effects of variations in suckling stimulus (changes in letter size) and exposure to high ambient temperatures on the quality and quantity of rat's milk and mammary tissue structure were determined. By increasing the number of suckling pups per dam, it was found that the amount of milk produced by the animals was proportionally increased, as each pup was able to drink the same amount of milk per day. With more than 6 pups per litter, the quality of the milk was not significanly changed. Smaller letters led to changes in the amount of milk suckled per milking and to considerable changes in the content of the milk. Exposure to high ambient temperatures, although for only 8 hr per day, drastically affected the amount of milk produced by the lactating rat as well as the quality of the milk produced. Microscopic examination of mammary tissue revealed changes in the alveolar area and height of the alveolar cells as the suckling stimulus varied. After heat exposure the gland changed to one of diminished synthesizing capability. It was concluded that 10-pup litters are the optimal litter size for research concerning rat milk, and the exposure to high ambient temperatures affects the milk yield by directly affecting the alveolae.     

Wood remains from Tel Nami,a middle bronze IIa and late bronze IIb port,local exploitation of trees and levantine cedar trade

Thirteen Middle Bronze Age IIa and four Late Bronze Age IIb (ca. 1950-1750 B.C. and thirteenth century, B.C., respectively)pieces of charcoal or water logged wood were found in the recent excavations of Tel Nami, a small port on the coast near Mount Carmel, Israel. These includedCedrus libani (cedar of Lebanon) (three samples), and local tree species that still grow today in the vicinity of the site—Pinus halepensis (Aleppo pine) (one sample),Olea europaea (olive tree) (five samples),Quercus calliprinos (kermes oak) (three samples),Quercus ithaburensis (Mt. Tabor oak) (four samples), andQuercus sp. (one sample). The discovery of Cedrus libani in a Middle Bronze Age IIa port is one of the earliest published examples of cedar wood from Israel. Together with other artifactual evidence for maritime trade from Tel Nami, this find suggests that a maritime trade in cedar wood existed along the Levantine coast.     

Repression of alkaline phosphatase in Salmonella typhimurium carrying a phoA+ phoR- episome from Escherichia coli.

Salmonella typhimurium does not produce alkaline phosphatase (nor beta-galactosidase). Nevertheless, it has the function of the phoR+ regulatory gene but lacks the function of the lacI+ regulatory gene. Several periplasmic proteins are derepressed when cells of S. typhimurium are starved for inorganic phosphate. The role of phoR is discussed.