High-affinity RNA targets of the Survival Motor Neuron protein reveal diverse preferences for sequence and structural motifs.
Nucleic Acids Res
; 46(20): 10983-11001, 2018 11 16.
Article
em En
| MEDLINE
| ID: mdl-30165668
ABSTRACT
The Survival Motor Neuron (SMN) protein is essential for survival of all animal cells. SMN harbors a nucleic acid-binding domain and plays an important role in RNA metabolism. However, the RNA-binding property of SMN is poorly understood. Here we employ iterative in vitro selection and chemical structure probing to identify sequence and structural motif(s) critical for RNA-SMN interactions. Our results reveal that motifs that drive RNA-SMN interactions are diverse and suggest that tight RNA-SMN interaction requires presence of multiple contact sites on the RNA molecule. We performed UV crosslinking and immunoprecipitation coupled with high-throughput sequencing (HITS-CLIP) to identify cellular RNA targets of SMN in neuronal SH-SY5Y cells. Results of HITS-CLIP identified a wide variety of targets, including mRNAs coding for ribosome biogenesis and cytoskeleton dynamics. We show critical determinants of ANXA2 mRNA for a direct SMN interaction in vitro. Our data confirms the ability of SMN to discriminate among close RNA sequences, and represent the first validation of a direct interaction of SMN with a cellular RNA target. Our findings suggest direct RNA-SMN interaction as a novel mechanism to initiate the cascade of events leading to the execution of SMN-specific functions.
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
RNA
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Proteína 1 de Sobrevivência do Neurônio Motor
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Motivos de Nucleotídeos
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Domínios Proteicos
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article