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이선지,이성백,Paola Ramirez,변유리,김주성,정용수,백광희,윤재승 한국유전학회 2012 Genes & Genomics Vol.34 No.2
The Drosophila melanogaster retinophilin (rtp) gene encoding the protein containing MORN (Membrane Occupation and Recognition Nexus) motifs that are known to interact with the plasma membrane has been identified to be expressed predominantly in adult eyes by several independent studies. Isolation and characterization of rtp mutant flies showed that the gene is involved in the phototransduction process by interacting with NINAC (neither inactivation nor afterpotential C) protein in adult photoreceptor cells. The gene was also reported to be involved in phagocytosis in embryos. We examined rtp gene expression during D. melanogaster development and in adult head tissues. The results showed that the gene is expressed at detectable levels only in adult photoreceptor cells but not in other developmental stages and other adult tissues,confirming its phototransduction functions. The RTP protein contains only 4 MORN motifs, whereas 8 MORN motifs are reported to be required for interactions with the plasma membrane. We found that RTP protein is present free in the cytosol and also is bound peripherally to the plasma membrane;this binding ability was found to be modulated by light. Our results suggest that the D. melanogaster RTP protein is a light-regulated peripheral membrane protein of photoreceptor cells.
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The seed sequence is necessary but insufficient for downregulation of target genes by miR-608
백광희,이광태,최영철,변유리,윤세나,정용수,윤재승 한국유전학회 2016 Genes & Genomics Vol.38 No.6
MicroRNAs are small, non-coding RNAs that inhibit gene expression posttranscriptionally through interaction with the 30 untranslated region (30 UTR) of target mRNAs. The most important factor for downregulation of target genes by miRNA is the ‘‘seed region,’’ which encompasses nucleotides 2–7 at the 50 end of the miRNA. In this study, sequence determinants for efficient downregulation of target genes were investigated by employing growth-suppressive miR-608 and miR-4651, which shares the seed sequence with miR-608. Cell growth experiments revealed that miR-4651 and miR-608-scram in which the seed sequences were mutated did not inhibit the growth of A549 cells in contrast to wild-type miR-608, which significantly inhibited cell growth. When similarity to miR-608 was increased by replacing sequences of miR- 4651 with that of miR-608, cell growth was gradually more inhibited. Similar results were obtained from a luciferase reporter assay using a reporter plasmid containing the 30 UTR of BCL2L1 and from western blot analysis of BCL2L1, CCND3, and phosphoinositide 3-kinase regulatory subunit 2. Moreover, microarray analyses revealed that overexpression of miR-4651 and miR-608-scram resulted in inefficient downregulation of target genes, and the number of downregulated genes was increased when transfected with MT-3 mimic, which differs from miR-608 by four nucleotides located in the central region. Together, our findings provide a basis for understanding the mechanism underlying target recognition and/or downregulation of target genes by miR-608 and indicate that in addition to seed sequence, central and 30 parts of miR-608 play an important role in mediating efficient downregulation of target genes.
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