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H A Thieringer

Publications and source records attributed to H A Thieringer.

3 recordsLinked to original sources

Cold shock and adaptation.

Adaptation to environmental stresses, such as temperature fluctuation, is essential for the survival of all living organisms. Cellular responses in both prokaryotes and eukaryotes to high temperature include the synthesis of a set of highly conserved proteins known as the heat shock proteins. In contrast to the heat shock response, adaptation to low temperatures has not been as extensively studied. However, a family of cold-inducible proteins is evident in prokaryotes. In addition, most organisms have developed adaptive mechanisms that alter both membrane fluidity and the protein translation machinery at low temperature. This review addresses the different adaptive mechanisms used by a variety of organisms with a focus on the molecular mechanisms of cold adaptation that have recently been identified during the cold shock response in Escherichia coli.

Adaptation, Physiological↗

Identification and developmental characterization of a novel Y-box protein from Drosophila melanogaster.

The Y-box proteins are a family of highly conserved nucleic acid binding proteins which are conserved from bacteria to human. In this report we have identified a new member of this family from Drosophila melanogaster. Degenerate-PCR was used to identify a conserved region within the highly conserved cold-shock domain (CSD) of Y-box proteins. Subsequently, the cDNA for this gene was sequenced, and the identified open reading frame was named ypsilon schachtel (yps). The expression pattern of yps indicates that this gene is expressed throughout development with the highest level of expression found in adult flies. In situ hybridization shows that the yps mRNA is maternally loaded into the egg cytoplasm. In addition, there appears to be expression of yps mRNA in mesodermal tissue during embryogenesis. YPS, while containing a conserved CSD, is novel in that it completely lacks the alternating acidic and basic regions found in the C-terminus of the other vertebrate eukaryotic Y-box proteins. The CSD of yps was purified and gel-shift analysis showed that this domain can interact with RNA. We predict that YPS would be an RNA-binding protein due to these results and the motifs which have been identified within the amino acid sequence.

Amino Acid Sequence↗

Antisense RNA-mediated inhibition of mouse hepatitis virus replication in L2 cells.

We have successfully used antisense RNA to inhibit replication of the mouse hepatitis virus (MHV) in a cell culture system. MHV is a single-stranded RNA virus of positive polarity. Mouse L2 cells were stably transfected with an antisense construct that targets regions of genes 5 and 6 of the virus. High levels of expression from this construct, which is under control of the human elongation factor 1 alpha promoter, were found. After infection of the antisense cell lines with MHV, replication of the virus was significantly reduced compared with control cells. In a viral plaque assay, smaller plaques were found in the antisense cell lines. In addition, up to a 92% inhibition in the number of viral particles produced in one antisense cell line could be seen. This inhibitory effect decreased at longer (> 16 hour) infection times. It was possible to both increase the amount of inhibition and prolong the inhibitory effect by reducing the multiplicity of infection. Our results suggest that antisense RNA may be an effective tool to slow down progression of MHV infection in mice.

Animals↗