Engineered domain swapping indicates context dependent functional role of RNA G-quadruplexes.
RNA domain swapping typically demonstrates conservation of the native function of the domain in a non-native context. In contrast, we employed RNA engineering to demonstrate deviation of G-quadruplex (GQ) function that is contingent upon its context dependent location, which is opposite to their native functional role. Known translation repressing RNA GQs were engineered into human VEGF IRES A replacing the endogenous GQ domain essential for translation. Alternatively, the translation inhibitory GQ motif within the 5'-UTR of MT3-MMP mRNA was replaced with two known GQ motifs that are essential for translation. The results indicate that the engineered GQ domains can adopt GQ structures in a foreign environment with a functional role reversal to accommodate the need of the endogenous swapped motifs. The observations establish the functionality and context dependent modularity of RNA GQ structures.
Northeast Ohio Regional Sewer District, Cuyahoga Heights, OH, 44125, USA.
Debmalya
D
Bhattacharyya
Department of Chemistry and Biochemistry, Kent State University, Kent, OH, 44242, USA.
Mark J
MJ
Morris
Department of Chemistry and Biochemistry, Kent State University, Kent, OH, 44242, USA.
Prakash
P
Kharel
Molecular, Cellular and Developmental Biology, Howard Hughes Medical Institute, Yale University, New Haven, CT, 06520-8103, USA.
Gayan
G
Mirihana Arachchilage
Department of Chemistry and Biochemistry, Kent State University, Kent, OH, 44242, USA.
Katy M
KM
Fedeli
Department of Chemistry and Biochemistry, Kent State University, Kent, OH, 44242, USA. Electronic address: sbasu@kent.edu.
Soumitra
S
Basu
D020121:5' Untranslated Regions / Q000235:genetics*; D054856:G-Quadruplexes; D005786:Gene Expression Regulation; D006801:Humans; D053513:Matrix Metalloproteinase 16 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010442:Peptide Chain Initiation, Translational; D014176:Protein Biosynthesis; D000072417:Protein Domains; D015202:Protein Engineering; D012313:RNA / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D020121:5' Untranslated Regions; C507102:MMP16 protein, human; D012333:RNA, Messenger; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D012313:RNA; D053513:Matrix Metalloproteinase 16
Domain swapping; G-quadruplex; Nucleic acid structure-function relationship; RNA engineering