Cloning and expression of a human kinesin heavy chain gene: interaction of the COOH-terminal domain with cytoplasmic microtubules in transfected CV-1 cells.

To understand the interactions between the microtubule-based motor protein kinesin and intracellular components, we have expressed the kinesin heavy chain and its different domains in CV-1 monkey kidney epithelial cells and examined their distributions by immunofluorescence microscopy. For this study, we cloned and sequenced cDNAs encoding a kinesin heavy chain from a human placental library. The human kinesin heavy chain exhibits a high level of sequence identity to the previously cloned invertebrate kinesin heavy chains; homologies between the COOH-terminal domain of human and invertebrate kinesins and the nonmotor domain of the Aspergillus kinesin-like protein bimC were also found. The gene encoding the human kinesin heavy chain also contains a small upstream open reading frame in a G-C rich 5' untranslated region, features that are associated with translational regulation in certain mRNAs. After transient expression in CV-1 cells, the kinesin heavy chain showed both a diffuse distribution and a filamentous staining pattern that coaligned with microtubules but not vimentin intermediate filaments. Altering the number and distribution of microtubules with taxol or nocodazole produced corresponding changes in the localization of the expressed kinesin heavy chain. The expressed NH2-terminal motor and the COOH-terminal tail domains, but not the alpha-helical coiled coil rod domain, also colocalized with microtubules. The finding that both the kinesin motor and tail domains can interact with cytoplasmic microtubules raises the possibility that kinesin could crossbridge and induce sliding between microtubules under certain circumstances

P-10 Time Spent Studying, on Social Media, and Spirituality: Nursing Students’ Perceptions

This presentation highlights findings from a multi-university study exploring students’ perceptions of studying, social media, and spirituality. Mixed methods were utilized, an online survey was emailed, with N=88 sample size. Over half (55%) have a personal daily devotional 4 – 7 days/week

Fission yeast 26S proteasome mutants are multi-drug resistant due to stabilization of the pap1 transcription factor

Here we report the result of a genetic screen for mutants resistant to the microtubule poison methyl benzimidazol-2-yl carbamate (MBC) that were also temperature sensitive for growth. In total the isolated mutants were distributed in ten complementation groups. Cloning experiments revealed that most of the mutants were in essential genes encoding various 26S proteasome subunits. We found that the proteasome mutants are multi-drug resistant due to stabilization of the stress-activated transcription factor Pap1. We show that the ubiquitylation and ultimately the degradation of Pap1 depend on the Rhp6/Ubc2 E2 ubiquitin conjugating enzyme and the Ubr1 E3 ubiquitin-protein ligase. Accordingly, mutants lacking Rhp6 or Ubr1 display drug-resistant phenotypes

PCR-Mediated Epitope Tagging of Genes in Yeast

Epitope tagging of genes is a powerful technique facilitating assays for gene function, determination of subcellular distribution of proteins, affinity purification, study of protein interaction with other proteins, DNA or RNA, and any other antibody-based approach in the absence of protein-specific antibodies. Here, we describe a one-step PCR-based strategy for insertion of epitope tags at the chromosomal locus. This method takes advantage of efficient homologous recombination in yeast. PCR amplified tags are directed to desired chromosomal loci with the help of primer-encoded flanking homologous sequences enabling selective epitope tagging of genes of interest

Contamination source review for Building E2370, Edgewood Area, Aberdeen Proving Ground, Maryland

The US Army Aberdeen Proving Ground (APG) commissioned Argonne National Laboratory (ANL) to conduct a contamination source review to identify and define areas of toxic or hazardous contaminants and to assess the physical condition and accessibility of APG buildings. The information obtained from this review may be used to assist the US Army in planning for the future use or disposition of the buildings. The contamination source review consisted of the following tasks: historical records search, physical inspection, photographic documentation, and geophysical investigation. This report provides the results of the contamination source review for Building E2370. Many of the APG facilities constructed between 1917 and the 1960s are no longer used because of obsolescence and their poor state of repair. Because many of these buildings were used for research, development, testing, and/or pilot-scale production of chemical warfare agents and other military substances, the potential exists for portions of the buildings to be contaminated with these substances, their degradation products, and other laboratory or industrial chemicals. These buildings and associated structures or appurtenances may contribute to environmental concerns at APG

From START to FINISH : the influence of osmotic stress on the cell cycle

Peer reviewedPublisher PD

December 1965

Massachusetts Turf and Lawn Grass Council Better Turf Through Research and Educatio