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Illegal Beings: Human Clones
Specificity and determinants of Sam68 RNA binding. Implications for the biological function of K homology do
Lin Q, Taylor SJ, Shalloway DJ Biol Chem. 1997 Oct 24;272(43):27274-80..
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853, USA.
Sam68, a specific target of the Src tyrosine kinase in mitosis, possesses features common to RNA-binding proteins, including a K homology (KH) domain. To elucidate its biological function, we first set out to identify RNA species that bound to Sam68 with high affinity using in vitro selection. From a degenerate 40-mer pool, 15 RNA sequences were selected that bound to Sam68 with Kd values of 12-140 nM. The highest affinity RNA sequences (Kd approximately 12-40 nM) contained a UAAA motif; mutation to UACA abolished binding to Sam68. Binding of the highest affinity ligand, G8-5, was assessed to explore the role of different regions of Sam68 in RNA binding. The KH domain alone did not bind G8-5, but a fragment containing the KH domain and a region of homology within the Sam68 subgroup of KH-containing proteins was sufficient for G8-5 binding. Deletion of the KH domain or mutation of KH domain residues analogous to loss-of-function mutations in the human Fragile X syndrome gene product and the Caenorhabditis elegans tumor suppressor protein Gld-1 abolished G8-5 binding. Our results establish that a KH domain-containing protein can bind RNA with specificity and high affinity and suggest that specific RNA binding is integral to the functions of some regulatory proteins in growth and development.
This Message is being posted for educational purposes, as well as for comment and criticism, by the visitors to the HumanCloning.org Foundation website (http://www.humancloning.org ).
Disclaimer: This abstract is being posted for educational purposes, as well as for comment and criticism, by the visitors to the Human Cloning Foundation website (www.HumanCloning.org ). This abstract is representative of a larger article that is indexed on Medline.
The Human Cloning Foundation was established February, 1988. .
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