Analysis of the 5′ end of the mouse Elavl1 ( mHuA) gene reveals a transcriptional regulatory element and evidence for conserved genomic organization

• Published in: Gene Vol. 242; no. 1; pp. 125 - 131
• Main Authors: King, P.H., Fuller, J.J., Nabors, L.B., Detloff, P.J.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 25-01-2000
• Subjects: Amino Acid Sequence; Animals; Base Sequence; c-ets protein; CCAAT/response element-binding protein; Cell Line; Cloning, Molecular; Conserved Sequence; DNA - chemistry; DNA - genetics; ELAV1 gene; Elav11 gene; ELAV2 gene; Elavl1 gene; elavl4 gene; ELVAL1 gene; Embryo, Mammalian - metabolism; Gene Expression; Gene Expression Regulation; Gene Expression Regulation, Developmental; Genomic cloning; HeLa Cells; HUA gene; Humans; Luciferases - genetics; Luciferases - metabolism; mHuA gene; mHuD gene; Mice; Molecular Sequence Data; Promoter analysis; Promoter Regions, Genetic - genetics; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Regulatory Sequences, Nucleic Acid - genetics; Restriction Mapping; RNA - genetics; RNA - metabolism; RNA mapping; RNA-binding protein; RNA-Binding Proteins - genetics; Sequence Analysis, DNA; Tissue Distribution; Transcription, Genetic; RNA mapping; Elav, embryonic lethal abnormal visual; GAPDH, glyceraldehyde-3 phosphate dehydrogenase; RNP, ribonucleoprotein consensus motif; RT–PCR, reverse transcription–polymerase chain reaction; Genomic cloning; UTR, untranslated region; AP1, activator protein 1; TS, transcription site; RNA-binding protein; ES cell, embryonic stem cell; nt, nucleotide(s); RRM, RNA recognition motif; cDNA, DNA complementary to RNA; CREB, c-AMP response element binding protein; BAC, bacterial artificial chromosome; aa, amino acid; Promoter analysis
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/S0378-1119(99)00537-5

mHuA (
Elavl1) belongs to a highly conserved family of genes encoding RNA-binding proteins and has been linked to cell growth and proliferation through its regulation of mRNA stability. Here, we use an RNase protection assay to demonstrate that the
mHuA transcript is relatively abundant in a range of mouse tissues, with the highest levels being found in lung and embryonic stem cells. We then cloned and mapped an 18
kb DNA fragment which encompasses the 5′ end of the
mHuA gene. The genomic organization in this region is similar to the neural-restricted family members,
Hel-N1 (
ELAVL2) and
mHuD (
Elavl4). The first exon is lengthy and untranslated, and the second exon, which includes the methionine start site, ends between the ribonucleoprotein motifs of the first RNA binding domain. Mapping of the
mHuA transcript by primer extension demonstrated three potential transcription-initiation sites which were detected consistently among different tissues and cell lines. Analysis of the sequence flanking these sites revealed the presence of transcriptional elements including TATA, CREB, c-ets, and AP1 sites. Transfection analysis of this promoter region using a luciferase-reporter-gene assay indicated strong transcriptional activity both in HeLa and in mouse macrophage (RAW) cells which is consistent with the ubiquitous expression pattern of
mHuA. Thus, while the genomic organization of
mHuA is similar to the neural-restricted members of the
Elav family, the promoter element differs substantially both by sequence analysis and transcriptional activity in non-neural cell types.