Nitrogen regulation of protein–protein interactions and transcript levels of GlnK PII regulator and AmtB ammonium transporter homologs in Archaea

Gene homologs of GlnK PII regulators and AmtB-type ammonium transporters are often paired on prokaryotic genomes, suggesting these proteins share an ancient functional relationship. Here, we demonstrate for the first time in Archaea that GlnK associates with AmtB in membrane fractions after ammonium shock, thus, providing a further insight into GlnK-AmtB as an ancient nitrogen sensor pair. For this work, Haloferax mediterranei was advanced for study through the generation of a pyrE2-based counterselection system that was used for targeted gene deletion and expression of Flag-tagged proteins from their native promoters. AmtB1-Flag was detected in membrane fractions of cells grown on nitrate and was found to coimmunoprecipitate with GlnK after ammonium shock. Thus, in analogy to bacteria, the archaeal GlnK PII may block the AmtB1 ammonium transporter under nitrogen-rich conditions. In addition to this regulated protein–protein interaction, the archaeal amtB-glnK gene pairs were found to be highly regulated by nitrogen availability with transcript levels high under conditions of nitrogen limitation and low during nitrogen excess. While transcript levels of glnK-amtB are similarly regulated by nitrogen availability in bacteria, transcriptional regulators of the bacterial glnK promoter including activation by the two-component signal transduction proteins NtrC (GlnG, NRI) and NtrB (GlnL, NRII) and sigma factor σN (σ54) are not conserved in archaea suggesting a novel mechanism of transcriptional control.

A two-step mechanism for epigenetic specification of centromere identity and function

The basic determinant of chromosome inheritance, the centromere, is specified in many eukaryotes by an epigenetic mark. Using gene targeting in human cells and fission yeast, chromatin containing the centromere-specific histone H3 variant CENP-A is demonstrated to be the epigenetic mark that acts through a two-step mechanism to identify, maintain and propagate centromere function indefinitely. Initially, centromere position is replicated and maintained by chromatin assembled with the centromere-targeting domain (CATD) of CENP-A substituted into H3. Subsequently, nucleation of kinetochore assembly onto CATD-containing chromatin is shown to require either the amino- or carboxy-terminal tail of CENP-A for recruitment of inner kinetochore proteins, including stabilizing CENP-B binding to human centromeres or direct recruitment of CENP-C, respectively.

Performance of historic stone building materials and systems : observations by architects, engineers and building conservators : excerpts from workshop discussions held on August 10, 1987, funded by the U.S. Environmental Protection Agency and the U.S. Department of the Interior, National Park Service, Cultural Resources Acid Rain Research Program.

"June 1988."

An evaluation of vacuum equipment for collection of asbestos waste /

"Contract no. 210-77-0053."

Development and validation of methods for sampling and analysis of workplace toxic substances /

Mode of access: Internet.

A preliminary investigation of the performance of men's safety-toe footwear /

Mode of access: Internet.

A recommended approach to recirculation of exhaust air /

Mode of access: Internet.

NIOSH manual of sampling data sheets.

Mode of access: Internet.

NIOSH manual of sampling data sheets : supplement to the 1977 edition.

Mode of access: Internet.

Engineering control of occupational safety and health hazards : recommendations for improving engineering practice, education and research : summary report of the Engineering Control Technology Workshop Technical Panel.

Mode of access: Internet.

Summary of proceedings and testimony from the Public Forum for the Development of a National Research Strategy for Neurological and Communicative Disorders /

"June 1, 1979."

National silicosis conference ... : Final report[s] of the committee[s].

List of references in parts 1 and 2.

Beryllium sampling methods: Comparison of two personal sample collection methods with the AEC sample collection method as used for one year in a beryllium production facility. /

Item 499-F-2