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Three different cDNAs encoding mouse D-factor/LIF receptor
M Tomida1, Y Yamamoto-Yamaguchi, M Hozumi
1Department of Chemotherapy, Saitama Cancer Center Research Institute, Inc.
Abstract:
Three cDNAs for mouse differentiation-stimulating factor (D-factor)/leukemia inhibitory factor (LIF) receptor were isolated from a cDNA library prepared from the liver of a pregnant mouse. A probe for screening was prepared by the RT-PCR method using human cDNA sequences as primers. The mouse D-factor receptor cDNA encoded 1,092 amino acids, which had a marked homology with the human counterpart and consisted of signal sequence, extracellular, transmembrane, and cytoplasmic domains. The WSXWS motif found in members of the cytokine receptor family was also present in the extracellular domain of the mouse D-factor receptor. A second form of cDNA that had a 501 bp insertion was isolated. The insertion introduced a stop codon so that the mRNA encoded the soluble receptor lacking transmembrane and intracellular domains. Because the insertion contained polyadenylation signals, two different sizes of mRNA encoding the soluble receptor were produced, depending on whether or not it utilized these signals. Transcripts utilizing these signals were 2.6-3 kb in size, and were very abundantly expressed in the liver. Transcripts that did not use these signals were longer than 5 kb and of similar size to the mRNA for the cellular receptor.
Insights
Researchers isolated mouse differentiation-stimulating factor (D-factor)/leukemia inhibitory factor (LIF) receptor cDNAs, identifying a soluble form. This soluble receptor, abundant in the liver, is produced via alternative mRNA processing.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Differentiation-stimulating factor (D-factor), also known as leukemia inhibitory factor (LIF), plays a crucial role in cellular differentiation.
- Understanding the receptor for D-factor/LIF is essential for elucidating its biological functions.
Purpose of the Study:
- To isolate and characterize cDNAs encoding the mouse differentiation-stimulating factor (D-factor)/leukemia inhibitory factor (LIF) receptor.
- To identify potential isoforms of the D-factor/LIF receptor, including soluble forms.
Main Methods:
- Isolation of mouse liver cDNAs using a probe derived from human sequences.
- RT-PCR (Reverse Transcription Polymerase Chain Reaction) for probe preparation.
- Sequence analysis to identify receptor domains and alternative splicing events.
Main Results:
- Three cDNAs for the mouse D-factor/LIF receptor were isolated, showing high homology to the human counterpart.
- A soluble receptor isoform was identified, resulting from a 501 bp insertion that introduces a premature stop codon.
- Alternative polyadenylation of the soluble receptor mRNA leads to two distinct transcript sizes (2.6-3 kb and >5 kb), with shorter transcripts being highly abundant in the liver.
Conclusions:
- The mouse D-factor/LIF receptor exists in at least two forms: a full-length cellular receptor and a soluble receptor.
- The soluble D-factor/LIF receptor is generated through alternative splicing and exhibits differential mRNA processing, with abundant expression in the liver.
- These findings provide insights into the molecular mechanisms regulating D-factor/LIF signaling and receptor availability.