Pentadecamer-binding proteins : Definition of two independent protein-binding sites needed for functional activity
Sigvardsson, Mikael LU ; Bemark, Mats LU
and Leanderson, Tomas
LU
(1995)
In Molecular and Cellular Biology
15(3).
p.1343-1352
- Abstract
The SP6 κ-promoter pentadecamer (pd) element was found to be unable to stimulate transcription when present in one copy as the only promoter element in a minimal promoter but showed weak stimulatory activity when present as a multimer (four copies). One copy of the pd element acted synergistically with an octamer element, but not with a SP1 site, to stimulate transcription. The effect was orientation dependent with regard to the pd element. Gel shift analysis showed that pd-binding proteins were expressed in transformed as well as nontransformed B lymphocytes, irregardless of their differentiation stage, and in HeLa cells. Two major complexes, binding to different sites within the pd element, were observed in gel shifts. A... (More)
The SP6 κ-promoter pentadecamer (pd) element was found to be unable to stimulate transcription when present in one copy as the only promoter element in a minimal promoter but showed weak stimulatory activity when present as a multimer (four copies). One copy of the pd element acted synergistically with an octamer element, but not with a SP1 site, to stimulate transcription. The effect was orientation dependent with regard to the pd element. Gel shift analysis showed that pd-binding proteins were expressed in transformed as well as nontransformed B lymphocytes, irregardless of their differentiation stage, and in HeLa cells. Two major complexes, binding to different sites within the pd element, were observed in gel shifts. A low-molecular-weight form dominated in resting cells, while a higher-molecular-weight form appeared after mitogenic stimulation. Southwestern analysis showed that the low-molecular-weight pd-binding protein had a molecular mass of 35 kDa, which was confirmed by fractionation by denaturating polyacrylamide gel electrophoresis and molecular sieving. The higher-molecular-weight complex was sensitive to detergent treatment, while the low-molecular-weight complex was not. Mutation analysis showed that the two pd-binding complexes interacted with distinct sites within the element and that dual occupancy was required for functional activity. The functional synergy between the pd element and the octamer was more pronounced in plasmacytomas than in B-cell lymphomas.
(Less)
- author
- Sigvardsson, Mikael
LU
; Bemark, Mats
LU
and Leanderson, Tomas
LU
- organization
- publishing date
- 1995-03
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Molecular and Cellular Biology
- volume
- 15
- issue
- 3
- pages
- 1343 - 1352
- publisher
- American Society for Microbiology
- external identifiers
-
- scopus:0028895738
- pmid:7862127
- ISSN
- 0270-7306
- DOI
- 10.1128/MCB.15.3.1343
- language
- English
- LU publication?
- yes
- id
- 42208c58-453d-455f-8613-d9ffd315947a
- date added to LUP
- 2023-12-06 17:15:13
- date last changed
- 2024-01-04 15:38:16
@article{42208c58-453d-455f-8613-d9ffd315947a,
abstract = {{<p>The SP6 κ-promoter pentadecamer (pd) element was found to be unable to stimulate transcription when present in one copy as the only promoter element in a minimal promoter but showed weak stimulatory activity when present as a multimer (four copies). One copy of the pd element acted synergistically with an octamer element, but not with a SP1 site, to stimulate transcription. The effect was orientation dependent with regard to the pd element. Gel shift analysis showed that pd-binding proteins were expressed in transformed as well as nontransformed B lymphocytes, irregardless of their differentiation stage, and in HeLa cells. Two major complexes, binding to different sites within the pd element, were observed in gel shifts. A low-molecular-weight form dominated in resting cells, while a higher-molecular-weight form appeared after mitogenic stimulation. Southwestern analysis showed that the low-molecular-weight pd-binding protein had a molecular mass of 35 kDa, which was confirmed by fractionation by denaturating polyacrylamide gel electrophoresis and molecular sieving. The higher-molecular-weight complex was sensitive to detergent treatment, while the low-molecular-weight complex was not. Mutation analysis showed that the two pd-binding complexes interacted with distinct sites within the element and that dual occupancy was required for functional activity. The functional synergy between the pd element and the octamer was more pronounced in plasmacytomas than in B-cell lymphomas.</p>}},
author = {{Sigvardsson, Mikael and Bemark, Mats and Leanderson, Tomas}},
issn = {{0270-7306}},
language = {{eng}},
number = {{3}},
pages = {{1343--1352}},
publisher = {{American Society for Microbiology}},
series = {{Molecular and Cellular Biology}},
title = {{Pentadecamer-binding proteins : Definition of two independent protein-binding sites needed for functional activity}},
url = {{http://dx.doi.org/10.1128/MCB.15.3.1343}},
doi = {{10.1128/MCB.15.3.1343}},
volume = {{15}},
year = {{1995}},
}