The Structure of Spliceosome
The standard spliceosome contains five small nuclear ribonuclear proteins, U1, U2,U4, U5, and U6, and several spliceosome associated proteins factors. Therefore, understanding this large complex machinery is a hard nut to crack for the scientists. But there has been significant progress in understanding and analyzing the structure of the spliceosomes through electron microscopy, crystallography, and other techniques. With the help of the advancements in the scientific techniques, the spliceosomes are identified as ribonuclear proteins as the complex contains both the RNA and proteins at its core machinery. A detailed examination of these complexes and their spatial arrangements proves that these spliceosomes are a kind of RNP enzyme, the snRNP, U1, and the similar proteins are named so because of their discovery and abundance. They are recognized as Uridine rich RNA molecules and numbered according to their availability. The canonical assembly of the spliceosomes consists of a no of elements that includes 5â€™ end splice site (5â€™SS), branch point sequence (BPS), 3â€™ splice site (3â€™ SS), and many other protein factors. Other factors can be up to 200 in number. That is why it is said to be a large complex with so many molecules involved, much of which is yet to be discovered.
The functioning of the Spliceosomes
The whole complex as an enzyme is not an intact molecule that acts on a particular reaction and goes on. Rather they are assembled stepwise as its working proceeds. The spliceosomes have to expel the introns from the precursor mRNA and the exons are to be attached during the process of splicing. For this, they have to get assembled by U1 recognizing the beginning of an intron at the 5â€™ end splice site (SS) and U2 recognizes the other end of the 3â€™ SS, so many rearrangements occur after and finally the cleavage of the introns and binding the two ends of the exons are exercised by the complex. The act of splicing is catalyzed by the enhancer and silencer sequences of nucleotides. The spliceosome assembly occurs through the interaction of the snRNPs and other protein factors in an orderly fashion. Once the assembly is completed, the process of splicing is catalyzed into two consecutive reactions. The first one includes the 2â€™OH group of the branch point adenosine upstream of the 3â€™ end of the intron reacts with the 5â€™ splice site with a novel phosphodiester bond between the 5â€™ end of the intron and the branch point. The second reaction comprises of the linkage of the 5â€™ and 3â€™ of the consecutive exons and the intron is excised out. The snRNP is again ready to carry out new splicing after its release from the completed cycle.
Effect of Splicing on Gene Expression
The presence of a vast number of genes for coding of the proteins, the splicing continues to be an essential process for the correct expression of the genes. The precursor mRNA has a lot of waste regions, not used for the coding of the specific protein. The splicing enables the correct coding by removing the introns and arranging the exons to be coded for the protein effectively. The spliceosomes not only remove the introns from the specific sequence but also affects the kinetics of the mRNA by correcting the sequence. It also enhances the translational efficiency in the cells.
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