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Interestingly, the progressive deletions of the integrin α10 promoter resulted in progressively diminished transcriptional activation, all the while not losing the activating effect of pRb up until the Δ55 construct suggesting a pRb responsive region between −108 bp to −55 bp upstream of the start of the site of transcription.
Furthermore, progressive deletions of the mouse UCP2 promoter (from -7.3 kb to +12 bp) indicated that an enhancer region (-86/-44) was responsible for both basal and FFA-stimulated UCP2 gene transcription.
To address the underlying mechanisms of these observations, we made progressive deletions of the MMP-2 cDNA from the 5' to the 3' end by PCR.
Fragments representing progressive deletions from both distal and proximal ends were amplified and cloned as EcoRI - BamHI fragments into pCaSpeR AUG βGal [39], or pPTGal [40].
We performed a structure-function analysis by testing progressive deletions of ORCTL3 for apoptosis induction.
A series of constructs were made with progressive deletions within EYFP (plasmids pPDF118, pPDF117, pPDF116 and pPDF115; Fig. 4A).
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By imaging the fluorescence recovery we found that the progressive deletion of the N-terminus of ERK1 caused an acceleration of nuclear trafficking (figure 4B).
Moreover we demonstrated that this difference is caused by a unique domain of ERK1 located at its N-terminus between residues 8 39, since the progressive deletion of this domain converts the shuttling features of ERK1 into those of ERK2.
This difference is caused by a domain of ERK1 located at its N-terminus since the progressive deletion of these residues converted the shuttling features of ERK1 into those of ERK2.
However, we cannot exclude that the progressive deletion of the C-terminus of Foxp3 leads to a loss of structural integrity per se, rather than invoking a requirement for cooperative binding of Runx1 and c-Rel.
In order to attempt to restrict expression to the lymphoid compartment, the pol II murine immunoglobulin heavy chain (IgH) intronic enhancer (Eμ) was placed at the 5' end of three constructs, with progressive deletion of the EBER1 promoter (Figure 1).
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