Common Variants of the Plant microRNA-168a Exhibit Differing Silencing Efficacy for Human Low-Density Lipoprotein Receptor Adaptor Protein 1 (LDLRAP1).
Microrna
; 8(2): 166-170, 2019.
Article
in En
| MEDLINE
| ID: mdl-30501607
ABSTRACT
BACKGROUND:
The discovery that a plant microRNA (miRNAs) from rice (Oryza sativa miR168a) can modify post-transcriptional expression of the mammalian. Low-Density Lipoprotein Receptor Adaptor Protein 1 (LDLRAP1) gene highlights the potential for cross-kingdom miRNAmRNA interactions.OBJECTIVE:
To investigate whether common variants of the conserved miR168a family have the capability for similar cross-kingdom regulatory functions, we selected sequences from three dietary plant sources rice (Oryza sativa), tomato (Solanum lycopersicum), apple (Malus domestica) and compared their ability to regulate human LDLRAP1 expression.METHODS:
Target prediction software intaRNA and RNAhybrid were used to analyze and calculate the energy and alignment score between the miR168a variants and human LDLRAP1 mRNA. An in vitro cell-based Dual-Luciferase® Reporter Assay (pmirGLO, Promega), was then used to validate the miRNA-mRNA interaction experimentally.RESULTS:
Computational analyses revealed that a single nucleotide difference at position 14 (from the 5' end of the miRNA) creates a GU wobble in the miRNA-mRNA duplex formed by tomato and apple miR168a variants. This GU wobble had only a small effect on the free energy score (-33.8-34.7 kcal/mol). However, despite reasonable hybridization energy scores (<-20 kcal/mol) for all miR168a variants, only the rice miR168a variant lacking a GU wobble significantly reduced LDLRAP1 transcript expression by 25.8 + 7.3% (p<0.05), as measured by relative luciferase activity.CONCLUSION:
In summary, single nucleotide differences at key positions can have a marked influence on regulatory function despite similar predicted energy scores and miRNA-mRNA duplex structures.Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Oryza
/
Solanum lycopersicum
/
Gene Expression Regulation, Plant
/
Malus
/
MicroRNAs
/
Adaptor Proteins, Signal Transducing
Type of study:
Prognostic_studies
Limits:
Humans
Language:
En
Journal:
Microrna
Year:
2019
Document type:
Article
Affiliation country:
New Zealand