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The main
focus of research in my lab is elucidating the structure and
function of catalytic core of the spliceosome, the multi-mega Dalton
ribonucleoprotein complex that performs splicing of mRNA precursors
in eukaryotes. We take advantage of a novel, minimal spliceosome we
have recently developed to learn about the function and evolution of
the spliceosome. This minimal system, which catalyzes a reaction
identical to the splicing reaction, consists of only two
spliceosomal snRNAs. In addition to providing direct evidence for
RNA catalysis in the spliceosome, and thus, settling the
longstanding and central question of the identity of the catalytic
domain, the minimal system provides a novel and powerful tool for
studying the spliceosome. Since the minimal system bypasses the
complicated spliceosomal assembly process, it offers the advantage
of a simple, functional system that is amenable to manipulations not
possible in the authentic spliceosome, and thus, provides the
long-sought possibility of functional analysis of the spliceosomal
catalytic core in isolation.
Currently
our studies on the minimal spliceosome follow three main lines of
research. First, we use the minimal splicing system as a simple
model of the catalytic domain of the spliceosome in order to
characterize the biochemistry and mechanistic details of the
splicing reaction. Next, the insights obtained from the minimal
spliceosome will be used as a basis for understanding how splicing
is performed in the authentic spliceosome. Specific conclusions
obtained from the minimal system will be tested in the authentic
spliceosome in order to obtain insight into the function of the
catalytic core within its physiological context. Importantly, the
minimal system provides a scaffold for analysis of the interactions
and function of spliceosomal protein and RNP factors. By adding
spliceosomal factors to the minimal system, we use biochemical
methods to obtain high-resolution structural and functional
information on their mode of interaction with the active site and
with each other, and their effect on catalysis. Taken together, in
addition to providing insight into the spliceosomal catalysis and
evolutionary origin, the above studies may ultimately make it
possible to build a more sophisticated minimal splicing system by
inclusion of suitable spliceosomal factors, which in turn will
provide a basis for further analysis of the organization and
function of the authentic spliceosome.
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