Subnuclear localization of the active variant surface glycoprotein gene expression site in Trypanosoma brucei

In Trypanosoma brucei, transcription by RNA polymerase II and 5′ capping of messenger RNA are uncoupled: a capped spliced leader is trans spliced to every RNA. This decoupling makes it possible to have protein-coding gene transcription driven by RNA polymerase I. Indeed, indirect evidence suggests that the genes for the major surface glycoproteins, variant surface glycoproteins (VSGs) in bloodstream-form trypanosomes, are transcribed by RNA polymerase I. In a single trypanosome, only one VSG expression site is maximally transcribed at any one time, and it has been speculated that transcription takes place at a unique site within the nucleus, perhaps in the nucleolus. We tested this by using fluorescence in situ hybridization. With probes that cover about 50 kb of the active 221 expression site, we detected nuclear transcripts of this site in a single fluorescent spot, which did not colocalize with the nucleolus. Analysis of marker gene-tagged active expression site DNA by fluorescent DNA in situ hybridization confirmed the absence of association with the nucleolus. Even an active expression site in which the promoter had been replaced by an rDNA promoter did not colocalize with the nulceolus. As expected, marker genes inserted in the rDNA array predominantly colocalize with the nucleolus, whereas the tubulin gene arrays do not. We conclude that transcription of the active VSG expression site does not take place in the nucleolus.

The microwave background anisotropies: Observations

Most cosmologists now believe that we live in an evolving universe that has been expanding and cooling since its origin about 15 billion years ago. Strong evidence for this standard cosmological model comes from studies of the cosmic microwave background radiation (CMBR), the remnant heat from the initial fireball. The CMBR spectrum is blackbody, as predicted from the hot Big Bang model before the discovery of the remnant radiation in 1964. In 1992 the cosmic background explorer (COBE) satellite finally detected the anisotropy of the radiation—fingerprints left by tiny temperature fluctuations in the initial bang. Careful design of the COBE satellite, and a bit of luck, allowed the 30 μK fluctuations in the CMBR temperature (2.73 K) to be pulled out of instrument noise and spurious foreground emissions. Further advances in detector technology and experiment design are allowing current CMBR experiments to search for predicted features in the anisotropy power spectrum at angular scales of 1° and smaller. If they exist, these features were formed at an important epoch in the evolution of the universe—the decoupling of matter and radiation at a temperature of about 4,000 K and a time about 300,000 years after the bang. CMBR anisotropy measurements probe directly some detailed physics of the early universe. Also, parameters of the cosmological model can be measured because the anisotropy power spectrum depends on constituent densities and the horizon scale at a known cosmological epoch. As sophisticated experiments on the ground and on balloons pursue these measurements, two CMBR anisotropy satellite missions are being prepared for launch early in the next century.