Related Experiment Videos
Characterization of an 11K protein produced by early region 3 of mouse adenovirus type 1
1Department of Genetics, University of Georgia, Athens 30602, USA.
Abstract:
Early region 3 (E3) of mouse adenovirus type 1 produces three mRNAs that can encode three proteins with unique carboxy-terminal exons. A bacterial fusion protein encoding the unique terminus of one the three predicted proteins was used to generate antiserum in rabbits. This antiserum detected a 14K protein on a Western blot of infected cell lysates. Immunoprecipitation and endoglycosidase H digestion revealed that the 14K protein was a glycoprotein with a core molecular weight of 11K, and we are designating this protein E3 gp 11K. Through in vitro translation experiments we determined that the previously predicted signal sequence of gp11K was cleaved when the protein was expressed during an infection. Biochemical and immunofluorescence microscopy data indicated that E3 gp11K was localized to the endoplasmic reticulum of infected cells. Biochemical experiments further indicated that gp11K is a peripheral membrane protein.
Insights
Mouse adenovirus type 1 E3 gp 11K is a glycoprotein localized to the endoplasmic reticulum. This protein, identified via specific antiserum, has a core molecular weight of 11K and is a peripheral membrane protein.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- Mouse adenovirus type 1 (MAdV-1) encodes proteins in its Early region 3 (E3).
- Three unique mRNAs are produced from the E3 region, each potentially encoding distinct proteins.
- Characterization of these E3-encoded proteins is crucial for understanding MAdV-1 infection.
Purpose of the Study:
- To identify and characterize a novel protein encoded by the E3 region of MAdV-1.
- To determine the molecular weight, post-translational modifications, and cellular localization of this E3 protein.
Main Methods:
- Generation of specific antiserum against a bacterial fusion protein representing a unique terminus of an E3 protein.
- Western blot analysis of infected cell lysates to detect the target protein.
- Immunoprecipitation and endoglycosidase H digestion to assess glycosylation and molecular weight.
- In vitro translation to confirm signal sequence cleavage.
- Immunofluorescence microscopy and biochemical assays to determine cellular localization and membrane association.
Main Results:
- A 14K protein, designated E3 gp 11K, was detected in MAdV-1 infected cells.
- E3 gp 11K is a glycoprotein with a core molecular weight of 11K.
- The signal sequence of E3 gp 11K is cleaved during infection.
- E3 gp 11K localizes to the endoplasmic reticulum.
- E3 gp 11K is a peripheral membrane protein.
Conclusions:
- The E3 region of MAdV-1 encodes a glycoprotein, E3 gp 11K.
- E3 gp 11K is processed and localized to the endoplasmic reticulum during MAdV-1 infection.
- This study provides initial characterization of a novel MAdV-1 E3 protein and its cellular location.