Both subtelomeric regions are required and sufficient for specific DNA fragmentation during macronuclear development in Stylonychia lemnae.

BACKGROUND: Programmed DNA-reorganization and DNA-elimination events take place frequently during cellular differentiation. An extreme form of such processes, involving DNA reorganization, DNA elimination and DNA fragmentation, is found during macronuclear differentiation in hypotrichous ciliates. Ciliated protozoa can therefore serve as a model system to analyze the molecular basis of these processes during cellular differentiation in eukaryotic cells. RESULTS: Using a biological approach to identify cis-acting sequences involved in DNA fragmentation, we show that in the hypotrichous ciliate Stylonychia lemnae sequences required for specific DNA processing are localized in the 3'- and the 5'-subtelomeric regions of the macronuclear precursor sequence. They can be present at various positions in the two subtelomeric regions, and an interaction between the two regions seems to occur. Sequence comparison revealed a consensus inverted repeat in both subtelomeric regions that is almost identical to the putative Euplotes chromosome breakage sequence (E-Cbs), also identified by sequence comparison. When this sequence was mutagenized, a processed product could no longer be detected, demonstrating that the sequence plays a crucial role in DNA processing. By injecting a construct into the developing macronucleus, which exclusively contains the subtelomeric regions of the Stylonychia alphal-tubulin gene, we show that subtelomeric regions are not only required but are also sufficient for DNA processing in Stylonychia. CONCLUSIONS: Our results indicate that an inverted repeat with the core sequence 5'-TGAA present in both subtelomeric regions acts as a Cbs in Stylonychia. The results allow us to propose a mechanistic model for DNA processing in this ciliate.

Cross section for b-Jet production in &pmacr;p collisions at radicals = 1.8 TeV

Bottom-quark production in &pmacr;p collisions at sqrt[s] = 1.8 TeV is studied with 5 pb(-1) of data collected in 1995 by the D0 detector at the Fermilab Tevatron Collider. The differential production cross section for b jets in the central rapidity region ( | y(b)|<1) as a function of jet transverse energy is extracted from a muon-tagged jet sample. Within experimental and theoretical uncertainties, D0 results are found to be higher than, but compatible with, next-to-leading-order QCD predictions.

Isolated photon cross section in p&pmacr; collisions at radicals = 1. 8 TeV

We report a new measurement of the cross section for the production of isolated photons with transverse energies ( E(gamma)(T)) above 10 GeV and pseudorapidities |eta|<2.5 in p&pmacr; collisions at sqrt[s] = 1.8 TeV. The results are based on a data sample of 107.6 pb(-1) recorded during 1992-1995 with the D0 detector at the Fermilab Tevatron collider. The background, predominantly from jets which fragment to neutral mesons, was estimated using the longitudinal shower shape of photon candidates in the calorimeter. The measured cross section is in good agreement with the next-to-leading order QCD calculation for E(gamma)(T) greater, similar36 GeV.

Differential production cross section of Z bosons as a function of transverse momentum at sqrt

We present a measurement of the transverse momentum distribution of Z bosons produced in p&pmacr; collisions at sqrt[s] = 1.8 TeV from data collected by the DO experiment at the Fermilab Tevatron Collider. We find good agreement between our results and current resummation calculations, and also use our data to extract nonperturbative parameters for a particular version of the resummation formalism. The resulting values are significantly more precise than obtained in previous determinations.

Search for second-generation leptoquark pairs in &pmacr;p collisions at radicals = 1.8 TeV

We have searched for second-generation leptoquark (LQ) pairs in the &mgr;&mgr;+jets channel using 94+/-5 pb(-1) of &pmacr;p collider data collected by the D0 experiment at the Fermilab Tevatron during 1993-1996. No evidence for a signal is observed. These results are combined with those from the &mgr;nu+jets and nunu+jets channels to obtain 95% confidence level (C.L.) upper limits on the LQ pair production cross section as a function of mass and beta, the branching fraction of a LQ decay into a charged lepton and a quark. Lower limits of 200(180) GeV/c(2) for beta = 1(1 / 2) are set at the 95% C.L. on the mass of scalar LQ. Mass limits are also set on vector leptoquarks as a function of beta.

Measurement of the W boson mass using electrons at large rapidities

We report a measurement of the W boson mass based on an integrated luminosity of 82 pb(-1) from p&pmacr; collisions at sqrt[s] = 1.8 TeV recorded in 1994-1995 by the D0 detector at the Fermilab Tevatron. We identify W bosons by their decays to enu, where the electron is detected in the forward calorimeters. We extract the mass by fitting the transverse mass and the electron and neutrino transverse momentum spectra of 11 089 W boson candidates. We measure M(W) = 80.691+/-0.227 GeV. By combining this measurement with our previously published central calorimeter results from data taken in 1992-1993 and 1994-1995, we obtain M(W) = 80.482+/-0.091 GeV.

Small-angle muon and bottom-quark production in p&pmacr; collisions at radicals = 1.8 TeV

This Letter describes a measurement of the muon cross section originating from b-quark decay in the forward rapidity range 2.4<| y(&mgr;)|<3.2 in p&pmacr; collisions at sqrt[s] = 1.8 TeV. The data used in this analysis were collected by the D0 experiment at the Fermilab Tevatron. We find that next-to-leading-order QCD calculations underestimate b-quark production by a factor of 4 in the forward rapidity region.

Spin correlation in t&tmacr; production from p&pmacr; collisions at radicals = 1.8 TeV

The D0 collaboration has performed a study of spin correlation in t&tmacr; production for the process t&tmacr;-->bW(+)&bmacr;W-, where the W bosons decay to enu or &mgr;nu. A sample of six events was collected during an exposure of the D0 detector to an integrated luminosity of approximately 125 pb(-1) of sqrt[s] = 1.8 TeV p&pmacr; collisions. The standard model (SM) predicts that the short lifetime of the top quark ensures the transmission of any spin information at production to the t&tmacr; decay products. The degree of spin correlation is characterized by a correlation coefficient kappa. We find that kappa>-0.25 at the 68% confidence level, in agreement with the SM prediction of kappa = 0.88.

Location of rDNA in the heteromeric macronucleus of Chilodonella steini.

As it was shown previously the chromatin of the heteromeric macronucleus of Chilodonella steini consists of two specific zones, the outer one (the orthomere) made up from late replicating electron dense material, and the inner zone (the paramere) of medium electron density and early replicating, with a concentration of electron dense material called the endosome in its central part. New details regarding the reorganization of the chromatin after macronuclear division are presented here. After the division no endosome is visible. It reappears within the first two hours after the separation of sister macronuclei. With the use of in situ rDNA - DNA hybridization it was shown that the rDNA in the macronucleus of Chilodonella steini is located in the outer zone (orthomere) and the endosome. Since it has been found earlier that the DNA content of the macronucleus depends on culture conditions the authors suggest that the quantitative DNA changes are caused preferentially by differential replication of rDNA dependent on the food conditions of the cells.

Recent advances in the study of ciliate genes.

This review gives a brief report on some recent advances in the study of molecular biology of ciliate genes. The main emphasis will be put on those results which may contribute to a better understanding of how gene structure and genome organization has evolved in ciliates. New data will be discussed in relation to the contribution they might give for a general understanding of eukaryotic gene evolution. The whole topic is divided into four parts: (1) a short introduction into genome organization of ciliates and an account of new insights in this field; (2) new results about telomere formation and replication; (3) regulation of transcription with new details about the atypical usage of the genetic code; (4) some speculations on unresolved problems and their possible solution by new methods. Other important new aspects of the molecular biology of ciliates which would fit well into this context, for example RNA self-splicing and evolutionary divergence of 17 S rRNA sequences, are not included as excellent newer reviews are available [e.g., 13-15, 42].

Characterization of macronuclear DNA in five species of ciliates.

Macronuclear DNA of four hypotrichous and one holotrichous ciliate species was characterized by biochemical techniques. The renaturation kinetics of the macronuclear DNAs of all five species were similar. Repetitive sequences occur only in an amount below 2%. Although the DNA content of the macronuclei of the species differs considerably, the kinetic complexity of the macronuclear DNA is rather uniform (around 3 x 10(10) daltons, i.e., 4-11 x the E. coli genome). Only in the macronuclei of the hypotrichous species the DNA exists as gene-sized fragments.

Histone genes in macronuclear DNA of the ciliate Stylonychia mytilus.

DNA in the macronucleus of Stylonychia mytilus exists as discrete gene-sized fragments which are derived from micronuclear DNA through a series of well-defined developmental events. It has been proposed that each of the DNA fragments might represent a gene and its controlling elements. We have investigated this possibility using genes which code for the five histone proteins. Macronuclear DNA fragments were fractionated according to size by agarose gel electrophoresis, the fragments transferred to nitrocellulose filters using the technique of Southern, and the filter-bound DNA hybridized with labeled cloned histone genes of the sea urchin, Psammechinus miliaris. Results indicate, first, that sequences homologous to the five individual histone gene probes are present in discrete macronuclear fragments which appear as bands in the gel hybridization assay. Secondly, for each of the five individual histone gene probes the homologous DNA fragments are several in number, ranging in size in from 7.6 Kb (Kilo base pairs) to 0.73 Kb. For example, the largest of six detected fragments hybridizing to the H3 gene probe contains approximately 10 times the amount of DNA required to code for a Stylonychia H3 histone. The smallest detected fragment hybridizing to the H3 probe contains enought DNA to code for approximately two copies of the histone. Finally, in general, no two histone approximately two copies of the histone. Finally, in general, no two histone gene probes hybridized to the same macronuclear DNA fragment. This result indicates that genes coding for the five histones in Stylonychia are not located together on the same macronuclear DNA fragments and implies that the five functionally related genes would not be transcribed together as a polycistronic unit.

Chromatin structure in the macronucleus of the ciliate Stylonychia mytilus.

Evidence is presented that macronuclear chromatin in the hypotrichous ciliate Stylonychia mytilus occurs in discrete fragments, each representing at least single genes. The size of these fragments varies between 3 and more than 70 nucleosomes with an average length of about 18 nucleosomes. This observation is discussed with respect to macronuclear structure of hypotrichous ciliates.

Free genes for rRNAs in the macronuclear genome of the ciliate Stylonychia mytilus.

When separated on an agarose gel, macronuclear DNA of the hypotrichous ciliate Stylonychia mytilus gives rise to many well-defined bands ranging in molecular weight from 0.3 x 10(6) to 14 x 10(6) dalton. Hybridization of 25 S rRNA, 17S rRNA or 5 S RNA to such a gel revealed sharp hybridization bands. This suggests that this banding pattern is not an artefact due to nonspecific degradation of macronuclear DNA but that the DNA in the macronucleus of Stylonychia occurs in discrete fragments, each coding for at least one gene. The size of the DNA fragment coding for rRNA was found to be 4.5 x 10(6) dalton, the fragment coding for 5 S RNA has a molecular weight of 150,000-250,000 dalton.