ABSTRACT
mHuA (Elavl1) belongs to a highly conserved family of genes encoding RNA-binding proteins and has been linked to cell growth and proliferation through its regulation of mRNA stability. Here, we use an RNase protection assay to demonstrate that the mHuA transcript is relatively abundant in a range of mouse tissues, with the highest levels being found in lung and embryonic stem cells. We then cloned and mapped an 18 kb DNA fragment which encompasses the 5' end of the mHuA gene. The genomic organization in this region is similar to the neural-restricted family members, Hel-N1 (ELAVL2) and mHuD (Elavl4). The first exon is lengthy and untranslated, and the second exon, which includes the methionine start site, ends between the ribonucleoprotein motifs of the first RNA binding domain. Mapping of the mHuA transcript by primer extension demonstrated three potential transcription-initiation sites which were detected consistently among different tissues and cell lines. Analysis of the sequence flanking these sites revealed the presence of transcriptional elements including TATA, CREB, c-ets, and AP1 sites. Transfection analysis of this promoter region using a luciferase-reporter-gene assay indicated strong transcriptional activity both in HeLa and in mouse macrophage (RAW) cells which is consistent with the ubiquitous expression pattern of mHuA. Thus, while the genomic organization of mHuA is similar to the neural-restricted members of the Elav family, the promoter element differs substantially both by sequence analysis and transcriptional activity in non-neural cell types.
Subject(s)
Promoter Regions, Genetic/genetics , RNA-Binding Proteins/genetics , Regulatory Sequences, Nucleic Acid/genetics , Amino Acid Sequence , Animals , Base Sequence , Cell Line , Cloning, Molecular , Conserved Sequence , DNA/chemistry , DNA/genetics , Embryo, Mammalian/metabolism , Gene Expression , Gene Expression Regulation , Gene Expression Regulation, Developmental , HeLa Cells , Humans , Luciferases/genetics , Luciferases/metabolism , Mice , Molecular Sequence Data , RNA/genetics , RNA/metabolism , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Restriction Mapping , Sequence Analysis, DNA , Tissue Distribution , Transcription, GeneticABSTRACT
Our goal was to estimate knee and hip joint contact forces during a variety of unconstrained "stretching" exercises that are often recommended for people with arthritis. The population of interest was aged females with and without significant osteoarthritis (OA). Three-dimensional (3-D) kinematic, force platform, and selected electromyographic (EMG) data were measured. A relatively standard and reasonably efficient technique was used for all subjects and tasks: inverse dynamics to estimate joint reaction forces and net moments, followed by heuristic reductionist techniques for predicting muscle load-sharing. Muscle force predictions were compatible with measured EMG activity for some tasks but less so for others, especially those with significant axial and medio-lateral movement components or high co-contraction. Such results suggest several improvements, which come at computational cost: 3-D joint models which better document muscle and passive tissue loading-sharing in abduction and axial torsion, and dynamic or novel static optimization approaches that can inherently predict muscle co-contraction. Nonetheless, the predicted joint contact loadings provide estimates that are essentially lower bounds on the likely joint contact loads. Based on our results we suggest that the side-kick and back-kick tasks be modified because the loading levels on the support leg are potentially excessive for the aged arthritic population.
