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A coding RNA segment that enhances the ribosomal recruitment of chicken ccn1 mRNA
Author(s) -
Mukudai Yoshiki,
Kubota Satoshi,
Eguchi Takanori,
Sumiyoshi Kumi,
Janune Danilo,
Kondo Seiji,
Shintani Satoru,
Takigawa Masaharu
Publication year - 2010
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.22894
Subject(s) - messenger rna , rna , open reading frame , microbiology and biotechnology , biology , translational frameshift , ribosomal rna , complementary dna , gene , coding region , ribosome , biochemistry , peptide sequence
CCN1, a member of the CCN family of proteins, plays important physiological or pathological roles in a variety of tissues. In the present study, we initially found a highly guanine–cytosine (GC)‐rich region of approximately 200 bp near the 5′‐end of the open reading frame, which was always truncated by amplification of the corresponding cDNA region through the conventional polymerase chain reaction. An RNA in vitro folding assay and selective ribonuclease digestion of the corresponding segment of the ccn1 mRNA confirmed the involvement of a stable secondary structure. Subsequent RNA electromobility‐shift assays demonstrated the specific binding of some cytoplasmic factor(s) in chicken embryo fibroblasts to the RNA segment. Moreover, the corresponding cDNA fragment strongly enhanced the expression of the reporter gene in cis at the 5′‐end, but did not do so at the 3′‐end. According to the results of a ribosomal assembly test, the effect of the mRNA segment can predominantly be ascribed to the enhancement of transport and/or entry of the mRNA into the ribosome. Finally, the minimal GC‐rich mRNA segment that was predicted and demonstrated to form a secondary structure was confirmed to be a functional regulatory element. Thus, we here uncover a novel dual‐functionality of the mRNA segment in the ccn1 open reading frame, which segment acts as a cis ‐element that mediates posttranscriptional gene regulation, while retaining the information for the amino acid sequence of the resultant protein. J. Cell. Biochem. 111: 1607–1618, 2010. © 2010 Wiley‐Liss, Inc.