DNA sequences of telomeres maintained in yeast

Telomeres, the ends of eukaryotic chromosomes, have long been recognized as specialized structures. Their stability compared with broken ends of chromosomes suggested that they have properties which protect them from fusion, degradation or recombination. Furthermore, a linear DNA molecule such as that of a eukaryotic chromosome must have a structure at its ends which allows its complete replication, as no known DNA polymerase can initiate synthesis without a primer. At the ends of the relatively short, multi-copy linear DNA molecules found naturally in the nuclei of several lower eukaryotes, there are simple tandemly repeated sequences with, in the cases analysed, a specific array of single-strand breaks, on both DNA strands, in the distal portion of the block of repeats. In general, however, direct analysis of chromosomal termini presents problems because of their very low abundance in nuclei. To circumvent this problem, we have previously cloned a chromosomal telomere of the yeast Saccharomyces cerevisiae on a linear DNA vector molecule. Here we show that yeast chromosomal telomeres terminate in a DNA sequence consisting of tandem irregular repeats of the general form C1-3A. The same repeat units are added to the ends of Tetrahymena telomeres, in an apparently non-template-directed manner, during their replication on linear plasmids in yeast. Such DNA addition may have a fundamental role in telomere replication.     

Structure, expression and divergence of T-cell receptor beta-chain variable regions

Analysis of three new T-cell receptor beta-chain variable regions together with those in the literature indicates that they have both remarkable similarities and differences with those of immunoglobulin. Less than 10 V regions appear to predominate in the thymus. V beta sequences are much more heterogeneous at the amino acid level than are immunoglobulin V regions and they appear to diverge between species much more quickly, apparently the result of additional hypervariable regions. Three of these putative new hypervariable regions lie outside of the classical immunoglobulin binding site, an indication that important interactions may be occurring in these regions with polymorphic MHC determinants.     

Accelerator mass spectrometry radiocarbon ages of amino acid extracts from Californian palaeoindian skeletons

A decade ago, aspartic acid racemization ages were determined for some skeletal remains found in California, near La Jolla, Del Mar and Sunnyvale, suggesting that people were present in North America during the Upper Pleistocene. These ages were obtained from the aspartic acid racemization rate, which was calibrated using a radiocarbon date of 17,150 +/- 1,470 yr BP determined for a skeleton found in Laguna Beach, California. These studies generated an intense controversy not only about the antiquity of human beings in the New World but also about the validity of racemization-based ages, and prompted efforts to date the finds by other means. Here we have used accelerator mass spectrometry (AMS) to determine the radiocarbon ages of the amino acid extracts used in the original racemization studies. Our studies indicate that some of the controversial Californian skeletons, which had been assigned to the Upper Pleistocene, are in fact Holocene.     

Different postsynaptic events in two types of retinal bipolar cell

The first synapse in the vertebrate visual system is made between the photoreceptors and the biopolar cells. Bioplar cells fall into two distinct classes according to whether the cell hyperpolarizes or depolarizes to small centred spots of light. Most evidence indicates that the light-induced hyperpolarization of the photoreceptprs suppresses transmitter release from the synaptic terminals, and it is probable that the differences between the two bipolar cell classes results from the different actions of the photoreceptor transmitter. In analysing the membrane potential fluctuations in both types of bipolar cell we find that the voltage noise spectra differ. It is to be expected that postsynaptic noise would be composed of the sum of noise generated in and transmitted from the cones and the noise arising from the statistical nature of synaptic transmission. We report here evidence for two such components in the voltage noise spectra recorded from each type of bipolar cell. The differences in the frequency distribution of the presumed transmitter-related components indicates that the transmitter generates events of longer duration in the depolarizing biopolar cells.     

The joining of V and J gene segments creates antibody diversity

The variable regions of mouse kappa (kappa) chains are coded for by multiple variable (V) gene segments and multiple joining (J) gene segments. The V kappa gene segments code for residues 1 to 95; the J kappa gene segments code for residues 96 to 108 (refs 1-3). This gene organisation is similar to that encoding the V lambda regions. Diversity in V kappa regions arises from several sources: (1) there are multiple germ-line V kappa gene segments and J kappa gene segments; (2) combinatorial joining of V kappa gene segments with different germline J kappa gene segments; and possibly, (3) somatic point mutation, as postulated for V lambda gene segments. Also, from a comparison of the number of germ-line J kappa gene segments and amino acid sequences, it has been suggested that J kappa region sequences may be determined by the way V kappa and J kappa gene segments are joined. This report supports this model by directly associating various J kappa sequences with given J kappa gene segments.     

Left-handed Z-DNA is induced by supercoiling in physiological ionic conditions

In physiological ionic conditions (200 mM NaCl), the (dC-dG)16 and (dC-dG)13 blocks in plasmid pRW751 are in a left-handed state when the negative superhelical density of the plasmid is greater than 0.972. As the salt concentration decreases or when (dmC-dG) sequences are present, less negative supercoiling is required to induce the right- to left-handed DNA transition. Furthermore, the single strand-specific nuclease, S1, recognizes and cleaves aberrant structural features at the junction between neighbouring right- and left-handed DNA regions.     

Amplification of immunoglobulin lambda constant genes in populations of wild mice

The lambda immunoglobulin light chain (Ig lambda) locus of BALB/c inbred mice consists of two variable region gene segments (V lambda)1-3, and four constant region gene segments (C lambda)1,2,4,5. Each C lambda gene segment is associated with a unique joining segment (J lambda)2,4-7, and they are organized in two paired units, J3C3-J1C1 and J2C2-J4C4 (refs 4, 8). Using cDNA probes specific for C lambda 1 and C lambda 2 (ref. 9) we have analysed the genomic organization of the C lambda gene segments in wild-derived and inbred strains of mice. Although Southern blots of the genomic DNA of inbred mice show a constant pattern of hybridization, wild-derived mice show a high degree of variation in the number, size and intensity of hybridizing fragments. We have now found that, per haploid genome, mice of a Mus musculus musculus stock isolated from Sladeckovce, Czechoslovakia (CzII) have at least 12 C lambda segments, and mice of a Mus musculus domesticus stock 'Centreville Lights' from Centreville, Maryland (CL) have at least 8 C lambda segments. There appears to have been relatively recent amplifications of the C lambda gene segments in wild mice.