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A small upstream open reading frame causes inhibition of human major vault protein expression from a ubiquitous mRNA
K Holzmann1, I Ambrosch, L Elbling
1Division of Cell Biology, Institute of Cancer Research, Vienna University, Austria.
Abstract:
Overexpression of the major vault protein (MVP) has been linked to a multidrug resistance (MDR) phenotype. We describe a ubiquitously expressed MVP mRNA splice variant (long (L)-MVP) differing from the regular isoform (short (S)-MVP) within the 5'-leader. Only L-MVP mRNA contains a small upstream open reading frame which was proven to inhibit in vitro and in vivo MVP expression in cis. L-MVP represented an almost constant portion of total MVP mRNA in diverse normal tissues, but was more variable in malignant cell types. MDR sublines with altered MVP expression displayed changed S-MVP/L-MVP ratios as compared to their drug-sensitive counterparts. Our results suggest alternative splicing as one mechanism for regulation of MVP expression.
Insights
Major vault protein (MVP) expression is regulated by alternative splicing. A long splice variant (L-MVP) inhibits MVP expression, impacting multidrug resistance (MDR) phenotypes in cancer cells.
Area of Science:
- Molecular Biology
- Cell Biology
- Cancer Research
Background:
- Overexpression of the major vault protein (MVP) is associated with multidrug resistance (MDR).
- MVP plays a role in cellular resistance to various drugs.
- Understanding MVP regulation is crucial for cancer therapy.
Purpose of the Study:
- To investigate the role of MVP mRNA splice variants in regulating MVP expression.
- To determine the impact of these variants on MDR phenotypes.
- To explore alternative splicing as a mechanism for MVP regulation.
Main Methods:
- Identification and characterization of MVP mRNA splice variants.
- Analysis of the 5'-leader sequence, including upstream open reading frames (uORFs).
- Comparison of splice variant ratios in normal and malignant tissues, and drug-sensitive vs. MDR cell lines.
Main Results:
- A ubiquitously expressed long (L)-MVP splice variant was identified, differing in the 5'-leader from the short (S)-MVP isoform.
- L-MVP mRNA contains a uORF that inhibits MVP expression both in vitro and in vivo.
- The ratio of S-MVP to L-MVP mRNA varied between normal and malignant tissues and was altered in MDR cell sublines.
Conclusions:
- Alternative splicing of MVP mRNA is a significant regulatory mechanism.
- The L-MVP variant, through its uORF, acts as a negative regulator of MVP expression.
- Altered S-MVP/L-MVP ratios may contribute to the development of multidrug resistance in cancer.