TY - JOUR
T1 - Immunological characterization of an early cytomegalovirus single-strand DNA-binding protein with similarities to the HSV major DNA-binding protein
AU - Anders, David G.
AU - Kidd, Jennifer R.
AU - Gibson, Wade
N1 - Funding Information:
We thank David Knipe for DNA clone pSG18 Sa/A to HSV-1 ICPB, and Ken Powell for antiserum to HSV-2 ICP11/12 . This work was aided by Public Health Service Grants Al 13718, At 22711, and At 19373 from the National Institute of Allergy and Infectious Diseases, and American Cancer Society Institutional Grant IN-11 X . D .G .A. was supported by Public Health Service Individual National Research Service Award A107293-01 from the National Institute of Allergy and Infectious Diseases .
Copyright:
Copyright 2015 Elsevier B.V., All rights reserved.
PY - 1987/12
Y1 - 1987/12
N2 - Monospecific polyclonal antisera were prepared against the 129-kDa, early, single-strand DNA-binding protein (DB129) of strain Colburn cytomegalovirus (CMV), and used to study its distribution in infected cells and its relatedness to a proposed human CMV (HCMV) counterpart (DB140). Indirect immunofluorescence of fixed, infected human fibroblasts showed DB129 to be localized within the intranuclear inclusions characteristic of replicating CMV. Treatment of infected cells with 50 to 100 μg phosphonoformic acid per milliliter resulted in the overproduction of DB129 and its accumulation within nuclei, both inside the inclusions and in surrounding areas of the nucleoplasm, whereas treatment with 500 μg/ml prevented inclusion formation, and DB129 was localized at discrete points throughout the infected-cell nuclei. The sera cross-reacted an estimated 10% with HCMV DB140 in an indirect immunoassay, and their use in immunofluorescence localized DB140 to the nuclear inclusions of HCMV-infected cells. Their immunological cross-reactivity, as well as their similar biochemical properties and intracellular distribution, support the likelihood that DB129 and DB140 are the protein products of homologous genes. The relationship of these proteins to the herpes simplex major DNA-binding protein is discussed.
AB - Monospecific polyclonal antisera were prepared against the 129-kDa, early, single-strand DNA-binding protein (DB129) of strain Colburn cytomegalovirus (CMV), and used to study its distribution in infected cells and its relatedness to a proposed human CMV (HCMV) counterpart (DB140). Indirect immunofluorescence of fixed, infected human fibroblasts showed DB129 to be localized within the intranuclear inclusions characteristic of replicating CMV. Treatment of infected cells with 50 to 100 μg phosphonoformic acid per milliliter resulted in the overproduction of DB129 and its accumulation within nuclei, both inside the inclusions and in surrounding areas of the nucleoplasm, whereas treatment with 500 μg/ml prevented inclusion formation, and DB129 was localized at discrete points throughout the infected-cell nuclei. The sera cross-reacted an estimated 10% with HCMV DB140 in an indirect immunoassay, and their use in immunofluorescence localized DB140 to the nuclear inclusions of HCMV-infected cells. Their immunological cross-reactivity, as well as their similar biochemical properties and intracellular distribution, support the likelihood that DB129 and DB140 are the protein products of homologous genes. The relationship of these proteins to the herpes simplex major DNA-binding protein is discussed.
UR - http://www.scopus.com/inward/record.url?scp=0023460873&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0023460873&partnerID=8YFLogxK
U2 - 10.1016/0042-6822(87)90154-1
DO - 10.1016/0042-6822(87)90154-1
M3 - Article
C2 - 2825423
AN - SCOPUS:0023460873
VL - 161
SP - 579
EP - 588
JO - Virology
JF - Virology
SN - 0042-6822
IS - 2
ER -