Functional dissection of AP-1 transcription factors using the Gal4 adaptor assay.
نویسندگان
چکیده
In the Gal4 adaptor assay, leucine zipper-containing proteins are tethered indirectly to the promoters of Gal4-responsive reporter genes via synthetic protein chimeras consisting of a Gal4 DNA-binding domain with an attached leucine zipper. Ternary complexes composed of the DNA binding site, the adaptor protein, and a leucine zipper factor can stimulate reporter gene activity, provided that the latter component possesses a transcriptional activation domain. This system is used to assay the transcriptional function and the interactions between various AP-1 factors.
منابع مشابه
Biochemical dissection of AP-1 recruitment onto Golgi membranes
Recruitment of the Golgi-specific AP-1 adaptor complex onto Golgi membranes is thought to be a prerequisite for clathrin coat assembly on the TGN. We have used an in vitro assay to examine the translocation of cytosolic AP-1 onto purified Golgi membranes. Association of AP-1 with the membranes required GTP or GTP analogues and was inhibited by the fungal metabolite, brefeldin A. In the presence...
متن کاملFunctional investigation of the BRCA1 Val1714Gly and Asp1733Gly variants by computational tools and yeast transcription activation assay
Mutations in the BRCA1 gene are known to be a major cause of hereditary breast cancer. However, characterizing the point mutationsassociated with cancer in BRCA1 is challenging because the functional impact of most of them is still unknown. Nowadays, a variety of methods are employed to identify cancer-associated mutations in BRCA1. This study is aimed to ass...
متن کاملFunctional dissection of the interactions of stonin 2 with the adaptor complex AP-2 and synaptotagmin.
Synaptic vesicle recycling is in part mediated by clathrin-mediated endocytosis. This process involves the coordinated assembly of clathrin and adaptor proteins and the concomitant selection of cargo proteins. Here, we demonstrate that the endocytotic protein stonin 2 localizes to axonal vesicle clusters through its micro-homology domain. Interaction of this domain with synaptotagmin I is suffi...
متن کاملFunctional Investigation of the Novel BRCA1variant (Glu1661Gly) byComputationalTools andYeastTranscription Activation Assay
Introduction: Mutations in the BRCA1 gene are major risk factors for breast and ovarian cancers. However, the relationship between some BRCA1 mutations and cancer risk remains largely unknown. Cancer risk predictions could be improved by evaluation of the impairment degree in the BRCA1 functions due to a specific mutation. This study aimed to assess the functional effect of a novel variant (Glu...
متن کاملFunctional Investigation of the Novel BRCA1variant (Glu1661Gly) byComputationalTools andYeastTranscription Activation Assay
Introduction: Mutations in the BRCA1 gene are major risk factors for breast and ovarian cancers. However, the relationship between some BRCA1 mutations and cancer risk remains largely unknown. Cancer risk predictions could be improved by evaluation of the impairment degree in the BRCA1 functions due to a specific mutation. This study aimed to assess the functional effect of a novel variant (Glu...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research
دوره 3 8 شماره
صفحات -
تاریخ انتشار 1992