ABSTRACT
We previously described two nuclear cofactors, TRRAP and TIP49, that have functional roles in Myc-mediated oncogenesis. We have now expanded the analysis of these Myc-associated cofactors to investigate their roles in apoptosis and cell proliferation. Although TRRAP and TIP49 are both essential for transformation, TIP49 modulates c-Myc-mediated apoptosis whereas disruption of TRRAP activity has no apparent effect on apoptosis. We extended our analysis of TIP49 to show that it also binds to the E2F1 transactivation domain and modulates both transforming and apoptotic activities. These results indicate that individual cofactors differentially potentiate c-Myc and E2F1 functions.
Subject(s)
Apoptosis/physiology , Carrier Proteins/metabolism , Cell Cycle Proteins , DNA Helicases , DNA-Binding Proteins , Nuclear Proteins/metabolism , Proto-Oncogene Proteins c-myc/genetics , Transcription Factors/genetics , ATPases Associated with Diverse Cellular Activities , Adaptor Proteins, Signal Transducing , Adenovirus E1A Proteins/genetics , Adenovirus E1A Proteins/metabolism , Animals , Carrier Proteins/genetics , Cell Cycle/genetics , Cell Division/genetics , Cell Transformation, Neoplastic/genetics , Cells, Cultured , E2F Transcription Factors , E2F1 Transcription Factor , Fibroblasts/pathology , Genes, Dominant , Humans , Nuclear Proteins/genetics , Proto-Oncogene Proteins c-myc/metabolism , Rats , Transcription Factors/metabolismABSTRACT
Mitochondrial DNA (mtDNA) analysis of forensic samples typically is performed when the quantity and quality of DNA are insufficient for nuclear DNA analysis or when maternal relatives may be the only reference source. Many of the steps required in the analytical process are both lengthy and labor intensive. Therefore, improvements in the process that reduce labor without compromising the quality of the data are desirable. The current procedure requires purification of the amplicons by centrifugal filtration after PCR and prior to cycle sequencing. Because this method requires several manipulations to perform, alternate cleanup procedures were investigated. These include the use of 1) Qiagen QlAquick PCR Purification columns, 2) Concert Rapid PCR Purification columns, and 3) ExoSAP-IT reagent. When the yield of purified amplicons, quality of the sequence profile, and ease of assay were evaluated, the use of ExoSAP-IT reagent for post-amplification purification was chosen to replace the filtration method.
