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Mr. Nitta has effectively expressed and purified GST-lamin A, full length LAP2a-His, and pRB large
pocket-His. Using these reagents Mr. Nitta has demonstrated through a GST-pull down assay followed
by Western analysis that the presence of LAP2a enhances the pRB-lamin A direct interaction. Mr.
Nitta has confirmed this finding using mammalian expressed full length pRB. He has begun to map
the binding regions of pRB, lamin A, and LAP2a by analyzing domain specific mutations of each
molecule using the GST-pull down assay. Additionally, Mr. Nitta already has in his possession,
surface domain and tumor derived mutations of pRB and laminopathy derived mutations of lamin A.
Therefore once Mr. Nitta determines the general binding regions using the domain specific mutations,
he will be prepared to determine the specific residues necessary for binding. In addition, he is
also poised to determine which disease derived mutations interefere with pRB-lamin A binding.
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