Analysis of glucocorticoid receptor subspecies binding to DNA-cellulose and isolated nuclei

Gruol, Donald J.; Wolfe, Kristal A.; Safarian, Sheila

doi:10.1016/0022-4731(89)90101-5

Cited by 1 publication

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“…We had also shown earlier that the peak A subspecies is the predominant DNA-binding form of the receptor (Gruol & Wolfe, 1989;Gruol et al, 1989c) and is, therefore, diagnostic of the conversion of receptor into a DNA-binding protein. The reduced production of peak A (relative to the TA-receptor) seen with RU 486 in Figure 5A,B correlates well with the lower capacity of the RU 486-receptor to bind DNA.…”

Section: Dna-cellulosesupporting

confidence: 66%

Transformation of glucocorticoid receptors bound to the antagonist RU 486: effects of alkaline phosphatase

Gruol¹,

Wolfe²

1990

Biochemistry

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RU 486 is a synthetic steroid that binds avidly to glucocorticoid receptors without promoting their transformation into activated transcription factors. A significant part of this behavior has been shown to be due to a failure of the RU 486 bound receptor to be efficiently released from a larger (sedimenting at 8-9 S) multimeric complex containing the 90-kDa heat shock protein. Our studies have found that in vitro at 15 degrees C the RU 486-receptor was slowly released from the 8-9S complex and converted into a DNA binding protein by a process that could be blocked by sodium fluoride. Moreover, this transition was significantly accelerated by treatment with alkaline phosphatase. High-resolution anion-exchange chromatography showed that the profile of receptor subspecies released from the 8-9S complex (in the absence of phosphatase treatment) was different for the RU 486 bound receptor when compared to the receptor occupied by the agonist triamcinolone acetonide. Production of the earliest eluting receptor form (peak A) was inhibited with RU 486. Peak A had previously been shown to be the predominant form of the receptor possessing a capacity to bind DNA. Treatment of the RU 486-receptor with alkaline phosphatase increased the formation of the peak A subspecies as well as the capacity of receptor to bind DNA-cellulose. Taken together, the results indicate that phosphorylation of the receptor or a tightly bound factor contributes to defining the capacity with which individual steroids can promote dissociation of the 8-9S complex and conversion of the glucocorticoid receptor into a DNA-binding protein.

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Section: Dna-cellulosesupporting

confidence: 66%