A long non-coding RNA transcribed from conserved non-coding sequences contributes to the mouse prolyl oligopeptidase gene activation

Abstract

Note: Nucleotide sequence data for two types (2.2 kb and 2.8 kb) of lncPrep+96kb reported in this paper are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB849012 and AB849013, respectively. SummaryProlyl oligopeptidase (POP) is a multifunctional protease which is involved in many physiological events, but its gene regulatory mechanism is poorly understood. To identify novel regulatory elements of the POP gene, we compared the genomic sequences at the mouse and human POP loci and found six conserved noncoding sequences (CNSs) at adjacent intergenic regions. From these CNSs, four long noncoding RNAs (lncRNAs) were transcribed and the expression pattern of one (lncPrep+96kb) was correlated with that of POP. lncPrep+96kb was transcribed as two forms due to the different transcriptional start sites and was localized at the nucleus and cytoplasm, although more was present at the nucleus. When we knocked down lncPrep+96kb in the primary ovarian granulosa cell and a hepatic cell line, the POP expression was decreased in both cells. In contrast, overexpression of lncPrep+96kb increased the POP expression only in the granulosa cell. Because lncPrep+96kb was upregulated with the same timing as POP in the hormone-treated ovary, this lncRNA could play a role in the POP gene activation in the granulosa cell.Moreover, a downstream region of the human POP gene was also transcribed. We propose a novel mechanism for the POP gene activation.Keywords: prolyl oligopeptidase/ long noncoding RNA/ conserved noncoding sequence/ granulosa cell/ skeletal muscle 4

A genomic region transcribed into a long noncoding RNA interacts with the Prss42/Tessp‐2 promoter in spermatocytes during mouse spermatogenesis, and its flanking sequences can function as enhancers

“…RACE analyses were performed as previously described (Kurihara et al, ; Matsubara et al, ). For 5′RACE, reverse transcription was performed with RT‐1 primer, and the first PCR was performed with GSP1 and the Abridged Anchor Primer.…”

Characterization of the human TCAM1P pseudogene and its activation by a potential dual promoter–enhancer: Comparison with a protein‐coding mouse orthologue

“…Total RNAs from the human liver and testis were purchased from Clontech Laboratories, Inc. (Mountain View, USA) and reverse transcription‐polymerase chain reaction (RT‐PCR) was done as previously described [17]. Primer sequences are shown in Table 1 .…”

“…5′RACE and 3′RACE were performed as previously described [7,17]. All the amplified products were subcloned into a pBluescript vector (Stratagene, La Jolla, CA) by the TA‐cloning method, and at least 10 subclones were sequenced for each product.…”

Mouse prolyl oligopeptidase plays a role in trophoblast stem cell differentiation into trophoblast giant cell and spongiotrophoblast

“…Total RNAs were prepared and qRT-PCR was performed as previously described [14,33]. All the data were normalized to Aip.…”