Advantages and Limitations of UV Cross-linking Analysis of Protein-RNA
Interactomes in Microbes
Abstract
RNA-binding proteins (RBPs) govern the lifespan of nearly all
transcripts and play key roles in adaptive responses in microbes. A
robust approach to examine protein-RNA interactions involves irradiating
cells with UV light to form covalent adducts between RBPs and their
cognate RNAs. Combined with RNA or protein purification, these
procedures can provide global RBP censuses or transcriptomic maps for
all target sequences of a single protein in living cells. Recent
development of novel methods has quickly populated the RBP landscape in
microorganisms. Here, we provide an overview of prominent UV
cross-linking techniques which have been applied to investigate RNA
interactomes in microbes. By assessing their advantages and caveats,
this technical evaluation intends to guide the selection of appropriate
methods and experimental design as well as to encourage the use of
complementary UV-dependent techniques to inspect RNA-binding activity.