TY - JOUR
T1 - Assessing the role of intrinsic disorder in RNA-binding protein function
T2 - hnRNP K as a case study
AU - Malaney, Prerna
AU - Benitez, Oscar
AU - Zhang, Xiaorui
AU - Post, Sean M.
N1 - Publisher Copyright:
© 2022 Elsevier Inc.
PY - 2022/12
Y1 - 2022/12
N2 - RNA-binding proteins (RBPs) typically bind to RNA in a sequence-specific manner, resulting in post-transcriptional gene regulation. While the various classes of RNA-binding domains are largely structured, flexible linkers are frequently observed between them. Emerging evidence suggests that these unstructured regions may help spatially position the RNA-binding domains allowing for RNA binding and/or may contribute directly to RNA association via certain sequence motifs contained within them. The importance of these unstructured regions is widely appreciated; however, understanding their contribution to RNA binding, protein stability, and function has been difficult to ascertain. Thus, it is crucial to have a set of rapid and economical assays that do not require specialized instrumentation to study their impact on RBP function. Herein, we discuss the use of plate-based and cell-based thermal shift assays to study the impact of the intrinsically disordered region on the function of a highly conserved RBP, hnRNP K.
AB - RNA-binding proteins (RBPs) typically bind to RNA in a sequence-specific manner, resulting in post-transcriptional gene regulation. While the various classes of RNA-binding domains are largely structured, flexible linkers are frequently observed between them. Emerging evidence suggests that these unstructured regions may help spatially position the RNA-binding domains allowing for RNA binding and/or may contribute directly to RNA association via certain sequence motifs contained within them. The importance of these unstructured regions is widely appreciated; however, understanding their contribution to RNA binding, protein stability, and function has been difficult to ascertain. Thus, it is crucial to have a set of rapid and economical assays that do not require specialized instrumentation to study their impact on RBP function. Herein, we discuss the use of plate-based and cell-based thermal shift assays to study the impact of the intrinsically disordered region on the function of a highly conserved RBP, hnRNP K.
KW - CETSA
KW - hnRNP K
KW - Intrinsic disorder
KW - Poly-C binding proteins
KW - Protein thermal shift
UR - http://www.scopus.com/inward/record.url?scp=85141337496&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85141337496&partnerID=8YFLogxK
U2 - 10.1016/j.ymeth.2022.10.009
DO - 10.1016/j.ymeth.2022.10.009
M3 - Article
C2 - 36334888
AN - SCOPUS:85141337496
SN - 1046-2023
VL - 208
SP - 59
EP - 65
JO - Methods
JF - Methods
ER -