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M. Dixit, B. J. Katz, B. K. Ambati; Identification of Flt1 Alternative Splicing Regulatory Factors. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4965.
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© ARVO (1962-2015); The Authors (2016-present)
To elucidate the mechanism of alternative splicing of Flt1 and identification of factors controlling variable levels of soluble and membrane bound Flt1.
Electrophoretic mobility shift assay was done by using approximately 500bp long 10 probes covering intron 13 and boundaries and nuclear protein extract from HMVEC cells. Region showing binding with nuclear protein were further used for designing smaller probes (100bp) for purification of bound factors using MACS technology. Bound proteins were identified by using Mass Spectrophotometry.
A total of 22 bound proteins were identified. Data search indicates that RBMX RNA binding motif protein, PTRF polymerase I and transcript release factor, and LOC729423 similar to heterogeneous nuclear ribonucleoprotein A1 might be involved in the alternative splicing regulatory mechanism.
Further characterization is needed to confirm the roles of RBMX RNA binding motif protein, PTRF polymerase I and transcript release factor, and LOC729423 similar to heterogeneous nuclear ribonucleoprotein A1 in Flt1 alternative splicing.
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