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The surf-4 gene encodes a novel 30 kDa integral membrane protein
1Eukaryotic Gene Organization and Expression Laboratory, Imperial Cancer Research Fund, London, UK.
Molecular Membrane Biology
|April 1, 1995
Summary
The surf-4 gene
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- The surfeit locus is a conserved gene cluster in vertebrates.
- The surf-4 gene is part of this locus and its sequence is conserved between mouse and human.
Purpose of the Study:
- To characterize the structure and localization of the mouse surf-4 protein.
- To investigate the topology and membrane association of surf-4.
Main Methods:
- In vitro translation of surf-4 cDNA.
- Salt and detergent extraction of microsomal membranes.
- Proteolysis protection assays.
- Generation of c-Myc tagged surf-4 chimeras.
- Immunofluorescence microscopy.
Main Results:
- Mouse surf-4 protein predicted to have seven transmembrane domains and an ER retrieval motif.
- In vitro translation produced a 30 kDa membrane-associated protein.
- Proteolysis protection indicated at least two membrane-spanning domains.
- Immunofluorescence showed cytoplasmic localization, not at the plasma membrane, consistent with the ER retrieval motif.
Conclusions:
- The surf-4 protein is a membrane protein with a topology consistent with its predicted structure.
- The ER retrieval motif directs surf-4 to the endoplasmic reticulum.
- A structural model for surf-4 is proposed.