Studies with chymotrypsin and RNAase showing a heterooligomeric structure of the glucocorticoid receptor complex from rat liver which is stabilized by a low molecular weight factor

Tymoczko, John L; Anderson, E. E.; Lee, James H.; Unger, Alison L.

doi:10.1016/0167-4889(86)90229-6

Cited by 5 publications

(3 citation statements)

References 38 publications

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“…In fact, there were usually two peaks, one at approx. 5 S and a shoulder at 4 S. We had previously reported that RNAase treatment of the 7-8 S receptor generates a 5 S form, whereas highsalt treatment yields the 4 S receptor (Tymoczko et al, 1986). The alteration in s value was not due simply to dilution, as shown in Fig.…”

Section: Resultsmentioning

confidence: 57%

“…This can be achieved by passing the 7-8 S receptor over a DEAE-cellulose column, which yields the 4 S receptor (Sakaue & Thompson, 1977;Tymoczko & Lee, 1985), or by treating the 7-8 S receptor with RNAase A, yielding an approx. 5 S form (Tymoczko & Phillips, 1984;Rossini, 1985;Tymoczko et al, 1986). Here we have presented evidence that DEAE-cellulose-purified 9-10 S receptor can be converted directly into the slowersedimenting 4-5 S forms by a factor that is eluted in the low-salt wash when the receptor preparation is chromatographed on DEAE-cellulose.…”

Section: Discussionmentioning

confidence: 99%

“…We have previously presented evidence that the 9-10 S form of the receptor can be converted into a low-salt 7-8 S form which is associated with RNA (Tymoczko & Phillips, 1983;Tymoczko et al, 1984). Although the 7-8 S RNAassociated receptor can bind to DNA several times more efficiently than the unactivated glucocorticoid-receptor complex, the removal of the RNA by RNAase leads to further enhancement of DNA binding as well as conversion into a 5 S form (Tymoczko et al, 1984(Tymoczko et al, , 1986Rossini, 1985). Whether RNA is associated with the glucocorticoid-receptor complex while it is in the RNAase-resistant 9-10 S form is not clear.…”

Section: Introductionmentioning

confidence: 99%

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Identification of a protein from rat liver cytosol that enhances activation of the glucocorticoid receptor

Tymoczko

Ahern

Unger

et al. 1988

Biochemical Journal

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We have identified a factor from rat liver cytosol that enhances the DNA-cellulose-binding ability of the glucocorticoid receptor and lowers the sedimentation value from 9-10 S to 4-5 S. Cytosol is prepared in the presence of molybdate, and unactivated receptor is isolated by chromatography on DEAE-cellulose in the presence of molybdate. This receptor sediments at 9-10 S and has little affinity for DNA. If the molybdate is removed and the receptor is incubated at 25 degrees C with the low-salt wash of the DEAE-cellulose column, DNA binding is enhanced by 50-600% relative to controls incubated with buffer only. In addition, the factor present in the low-salt wash converts the 9-10 S receptor into a mixture of 5 S and 4 S forms. The factor must be present during the incubation in order to exert its maximal effect. Factor added after the incubation has only marginal effects on the DNA-binding ability of the receptor, indicating that the factor does not increase the DNA-binding ability of activated receptor. Moreover, the factor is significantly less effective on receptor that has been activated before incubation with the factor. These results suggest that the factor acts as an activation enhancer. Preliminary characterization indicates that the activation enhancer is a trypsin-sensitive protein of approx. 70,000 Da, whose activation-enhancing properties are inhibited by ATP. RNAase A, which has effects similar to those described above on the 7-8 S receptor, does not mimic the effects of the activation enhancer on the 9-10 S receptor.

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Section: Resultsmentioning

confidence: 57%