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Henderson, J. O.

Publications and source records attributed to Henderson, J. O..

2 recordsLinked to original sources

Analysis of Escherichia coli Flagellin Gene Expression in Simulated Spaceflight Growth Conditions

Population growth curves of Escherichia coli (E. coli) cultures propagated in spaceflight environments demonstrate an extended log phase duration and an increased final cell concentration compared to E. coli cultures grown on Earth. We suggest that during spaceflight, a lack of convective mixing in the growth medium alters a cells microenvironment such that the cell at first devotes its resources primarily to motility and later shifts most of its resources to reproduction. As flagellin is the major protein component of flagella, the motility structure of E. coli, we propose that the expression of the flagellin gene (fliC) can be used as a diagnostic marker for determining if the bacterium is in fact altering its allocation of cellular resources to motility during specific growth phases. We have used semiquantitative RT-PCR to analyze the expression of fliC in cultures grown in laboratory apparatus that simulate various aspects of the spaceflight environment. The data from this pilot study indicate that fliC expression increases initially under stationary, slow clinorotation, and fast clinorotation growth conditions, peaking at mid-exponential phase and decreasing dramatically in stationary phase. These data support the hypothesis that metabolic resources are being directed towards forming the flagellum, the bacteria motile structure, before shifting resources to growth, reproduction, and maintenance.

microbiology

Molecular Genetic Analysis of Rbm45/Drbp1: Genomic Structure, Expression, and Evolution

RNA recognition motif-type RNA-binding domain containing proteins (RBDPs) participate in RNA metabolism including regulating mRNA stability, nuclear-cytoplasmic shuttling, and splicing. Rbm45 is an RBDP first cloned from rat brain and expressed spatiotemporally during rat neural development. More recently, RBM45 has been associated with pathological aggregates in the human neurological disorders amyotrophic lateral sclerosis, frontotemporal lobar degeneration, and Alzheimers. Rbm45 and the neural developmental protein musashi-1 are in the same family of RDBPs and have similar expression patterns. In contrast to Musashi-1, which is upregulated during colorectal carcinogenesis, we found no association of RBM45 overexpression in human colon cancer tissue. In order to begin characterizing RNA-binding partners of Rbm45, we have successfully cloned and expressed human RBM45 in an Intein fusion-protein expression system. Furthermore, to gain a better understanding of the molecular genetics and evolution of Rbm45, we used an in silico approach to analyze the gene structure of the human and mouse Rbm45 homologues and explored the evolutionary conservation of Rbm45 in metazoans. Human RBM45 and mouse Rbm45 span {bsim}17 kb and 13 kb, respectively, and contain 10 exons, one of which is non-coding. Both genes have TATA-less promoters with an initiator and a GC-rich element. Downstream of exon 10, both homologues have canonical polyadenylation signals and an embryonic cytoplasmic polyadenylation element. Moreover, our data indicate Rbm45 is conserved across all metazoan taxa from sponges (phylum Porifera) to humans (phylum Chordata), portending a fundamental role in metazoan development.

genetics