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Gel shift experiments with fragments of the Trypanosoma brucei RNA-binding protein RBP10 by Bin Liu; Christine Clayton is a Medicine article available to read on EtoBox.

What is Gel shift experiments with fragments of the Trypanosoma brucei RNA-binding protein RBP10 about?

## Abstract ## Objective Trypanosoma brucei is a parasite of mammals and Tsetse flies, and control of mRNA stability is critical for parasite survival in the two different hosts. T. brucei RBP10 is a protein with a single RNA Recognition Motif (RRM) which is expressed only in the mammalian (bloodstream) form. Numerous observations suggest that RBP10 binds to procyclic-specific mRNAs and targets them for destruction, and there is also some evidence for selective binding of RBP10 to RNAs containing the motif UA(U)~6~. We here investigated this binding further. ## Results We tested in vitro binding of RBP10 to two different probes in solution. One contained two copies of UA(U)~6~, and the other two copies of a mutant version, UACUCUCU, which is inactive in regulation. An N-terminal segment of RBP10, including the RRM domain and 90 residues to its C-terminus, could be produced as soluble protein. This could bind both probes in vitro with similar affinities in the low micromolar range, which is not atypical for a single RRM. Soluble RBP10 therefore did not distinguish between UA(U)~6~ and UACUCUCU. Since no other sequences were tested, the requirements for RBP10 RNA binding remain to be

Who reads Gel shift experiments with fragments of the Trypanosoma brucei RNA-binding protein RBP10?

Author

Bin Liu; Christine Clayton

Publisher

Springer Science and Business Media LLC

Published

2022

Language

EN

Field

Medicine (Health Sciences)