26] Probing ribosome structure and function using short oligodeoxyribonucleotides

Abstract

To probe the function of these regions, small complementary oligo deoxyribonucleotides (cDNA probes) have been used to hybridize specific regions of the ribosomal RNA in the ribosomal subunits after which various functions of the ribosome are checked with the probe in place. The methods used to perform these studies will be discussed in this chapter. This chapter presents various specific applications such as subunit association sites, tRNA binding sites, and other sites. That certain rRNA regions may be involved in subunit association has been suggested by several different studies. Recent evidence suggested that the 807–809 region of the 23S rRNA might base-pair with the CCA 3´-terminus of tRNA. There are many other assays that can be used to determine the function of various regions of the rRNA. Clearly, binding of mRNA can be assayed as it comes in juxtaposition with the ribosome. All in all it appears that this technique offers a way of providing substantive insight into the structure and function of regions of rRNA in the ribosome.