Jw. Voss et al., AN ALTERNATIVE PIT-1 RNA SPLICING PRODUCT REVEALS MODULAR BINDING ANDNONMODULAR TRANSCRIPTIONAL ACTIVITIES OF THE POU-SPECIFIC DOMAIN, Molecular endocrinology, 7(12), 1993, pp. 1551-1560
We have identified a form of the pituitary-specific POU protein Pit-1
that results from deletion of the POU-specific (POUs) domain by altern
ative RNA splicing. This natural variant of Pit-1 (called Delta 4Pit-1
) has revealed several aspects of the function of the POU, domain. The
Delta 4Pit-1 protein was characterized using a Delta 4Pit-1-specific
antiserum. Further, selection assays of random oligonucleotide pools i
dentified binding site preferences for both wild type and Delta 4Pit-1
. Methylation interference, copper phenanthrolene, and missing contact
analyses were used to compare the binding characteristics of the two
forms of Pit-1 on a selected site. DNA binding affinity assays on seve
ral DNA elements revealed that the POU, domain contains a modular DNA
binding activity affecting the DNA binding affinity of the entire POU
domain on some, but not on other, DNA sites. Functional analysis on su
ch DNA elements has revealed that the POUs domain is an essential, but
nonmodular, component of the Pit-1 trans-activation domain dependent
on its natural context within the Pit-1 protein.