Determination of Aryl Hydrocarbon Receptor Nuclear Translocator Protein Concentration and Subcellular Localization in Hepatic and Nonhepatic Cell Culture Lines: Development of Quantitative Western Blotting Protocols for Calculation of Aryl Hydrocarbon Receptor and Aryl Hydrocarbon Receptor Nuclear Translocator Protein in Total Cell Lysates

1997 ◽  
Vol 52 (2) ◽  
pp. 202-211 ◽  
Author(s):  
Jennifer L. Holmes ◽  
Richard S. Pollenz
Keyword(s):  
Cell Culture ◽  
Subcellular Localization ◽  
Aryl Hydrocarbon Receptor ◽  
Western Blotting ◽  
Protein Concentration ◽  
Total Cell ◽  
Translocator Protein ◽  
Aryl Hydrocarbon ◽  
Quantitative Western Blotting

Determination of Copy Number of c-Myc Protein per Cell by Quantitative Western Blotting

1999 ◽  
Vol 269 (1) ◽  
pp. 66-71 ◽  
Author(s):  
Claudia Rudolph ◽  
Gerold Adam ◽  
Andreas Simm
Keyword(s):  
Western Blotting ◽  
Copy Number ◽  
Quantitative Western Blotting

Identification of functional domains of the aryl hydrocarbon receptor nuclear translocator protein (ARNT)

1994 ◽  
Vol 14 (9) ◽  
pp. 6075-6086
Author(s):  
S Reisz-Porszasz ◽  
M R Probst ◽  
B N Fukunaga ◽  
O Hankinson
Keyword(s):  
Aryl Hydrocarbon Receptor ◽  
Mutant Cell ◽  
Translocator Protein ◽  
Functional Domains ◽  
Recognition Sequence ◽  
Dimerization Domain ◽  
Alpha Helices ◽  
Helix Loop Helix ◽  
Aryl Hydrocarbon

The activated aryl hydrocarbon receptor (AHR) and the AHR nuclear translocator (ARNT) bind DNA as a heterodimer. Both proteins represent a novel class of basic helix-loop-helix (bHLH)-containing transcription factors in that (i) activation of AHR requires the binding of ligand (e.g., 2,3,7,8-tetrachlorodibenzo-p-dioxin [TCDD]), (ii) the xenobiotic responsive element (XRE) recognized by the AHR/ARNT heterodimer differs from the recognition sequence for nearly all other bHLH proteins, and (iii) both proteins contain a PAS homology region, which in the Drosophila PER and SIM proteins functions as a dimerization domain. A cDNA for mouse ARNT has been cloned, and potential functional domains of ARNT were investigated by deletion analysis. A mutant lacking all regions of ARNT other than the bHLH and PAS regions is unimpaired in TCDD-dependent dimerization and subsequent XRE binding and only modestly reduced in ability to complement an ARNT-deficient mutant cell line, c4, in vivo. Both the first and second alpha helices of the bHLH region are required for dimerization. The basic region is required for XRE binding but not for dimerization. Deletion of either the A or B segments of the PAS region slightly affects TCDD-induced heterodimerization, while deletion of the complete PAS region severely affects (but does not eliminate) dimerization. Thus, ARNT possesses multiple domains required for maximal heterodimerization. Mutants deleted for PAS A, PAS B, and the complete PAS region all retain some degree of XRE binding, yet none can rescue the c4 mutant. Therefore, both the PAS A and PAS B segments, besides contributing to dimerization, apparently fulfill additional, unknown functions required for biological activity of ARNT.

scholarly journals Identification of functional domains of the aryl hydrocarbon receptor nuclear translocator protein (ARNT).

1994 ◽  
Vol 14 (9) ◽  
pp. 6075-6086 ◽  
Author(s):  
S Reisz-Porszasz ◽  
M R Probst ◽  
B N Fukunaga ◽  
O Hankinson
Keyword(s):  
Aryl Hydrocarbon Receptor ◽  
Mutant Cell ◽  
Translocator Protein ◽  
Functional Domains ◽  
Recognition Sequence ◽  
Dimerization Domain ◽  
Alpha Helices ◽  
Helix Loop Helix ◽  
Aryl Hydrocarbon

The activated aryl hydrocarbon receptor (AHR) and the AHR nuclear translocator (ARNT) bind DNA as a heterodimer. Both proteins represent a novel class of basic helix-loop-helix (bHLH)-containing transcription factors in that (i) activation of AHR requires the binding of ligand (e.g., 2,3,7,8-tetrachlorodibenzo-p-dioxin [TCDD]), (ii) the xenobiotic responsive element (XRE) recognized by the AHR/ARNT heterodimer differs from the recognition sequence for nearly all other bHLH proteins, and (iii) both proteins contain a PAS homology region, which in the Drosophila PER and SIM proteins functions as a dimerization domain. A cDNA for mouse ARNT has been cloned, and potential functional domains of ARNT were investigated by deletion analysis. A mutant lacking all regions of ARNT other than the bHLH and PAS regions is unimpaired in TCDD-dependent dimerization and subsequent XRE binding and only modestly reduced in ability to complement an ARNT-deficient mutant cell line, c4, in vivo. Both the first and second alpha helices of the bHLH region are required for dimerization. The basic region is required for XRE binding but not for dimerization. Deletion of either the A or B segments of the PAS region slightly affects TCDD-induced heterodimerization, while deletion of the complete PAS region severely affects (but does not eliminate) dimerization. Thus, ARNT possesses multiple domains required for maximal heterodimerization. Mutants deleted for PAS A, PAS B, and the complete PAS region all retain some degree of XRE binding, yet none can rescue the c4 mutant. Therefore, both the PAS A and PAS B segments, besides contributing to dimerization, apparently fulfill additional, unknown functions required for biological activity of ARNT.

Regulation of Subcellular Localization of the Aryl Hydrocarbon Receptor (AhR)

2001 ◽  
Vol 389 (2) ◽  
pp. 207-217 ◽  
Author(s):  
Catherine A. Richter ◽  
Donald E. Tillitt ◽  
Mark Hannink
Keyword(s):  
Subcellular Localization ◽  
Aryl Hydrocarbon Receptor ◽  
Aryl Hydrocarbon
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