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PMID: 3977852 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The use of monoclonal antibodies to study the proteins specified by the transforming region of human adenoviruses.

The Biochemical journal ·Vol. 225 ·No. 3 ·1985-02-01 ·Pages 649-55

Blair Zajdel ME, Barker MD, Dixon SC, Blair GE

Abstract

Monoclonal antibodies against two of the proteins specified by one of the transforming genes (early region 1B) of human adenovirus type 2 have been produced and characterized. Two clones (RA1 and PA6), generated by fusion of mouse myeloma NSO cells with splenocytes from rats immunized with whole-cell lysates of an adenovirus-transformed rat cell line (F19), secreted antibodies against a 58 kDa protein. Another clone (DC1) produced antibodies against the same protein, and resulted from fusion of immune rat splenocytes with the rat myeloma Y3.Ag.1.2.3. Immunoprecipitation studies showed that all three antibodies recognized [35S]-methionine-labelled 58 kDa protein, and phosphorylated derivatives of the 58 kDa protein labelled with [32P]orthophosphate present in infected human cells. One clone (EC3) produced antibody against a 19 kDa protein also encoded by early region 1B, but not sharing sequence homology with 58 kDa. The identity of the 19 kDa protein recognized by the EC3 antibody was established by immunoprecipitation from lysates of labelled-infected cells and from products of cell-free translation directed by mRNA isolated from adenovirus 2-infected cells. Indirect immunofluorescent-antibody staining of infected human cells using the RA1 and EC3 antibodies revealed a nuclear location of the 58 kDa protein and a mainly cytoplasmic location of the 19 kDa protein.

MeSH Terms
Adenoviruses, Human/metabolism Antibodies, Monoclonal Cell Transformation, Viral Chemical Precipitation Methionine/metabolism Microscopy, Fluorescence Phosphates/metabolism Subcellular Fractions Viral Proteins/immunology,metabolism
Chemicals
Antibodies, Monoclonal Phosphates Viral Proteins Methionine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Blair Zajdel M E
Barker M D
Dixon S C
Blair G E
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26 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2250649
Published
1985-02-01
Pages
649-55
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1144640
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2250649 PMC Full Text View journal details
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