Structural and dynamical characterization of the Miz-1 zinc fingers 5-8 by solution-state NMR.

Myc-interacting zinc finger protein-1 (Miz-1) is a BTB/POZ transcription factor that activates the transcription of cytostatic genes, such as p15(INK4B) or p21(CIP1). The C-terminus of Miz-1 contains 13 consensus C2H2 zinc finger domains (ZF). ZFs 1-4 have been shown to interact with SMAD3/4, while the remaining ZFs are expected to bind the promoters of target genes. We have noted unusual features in ZF 5 and the linker between ZFs 5 and 6. Indeed, a glutamate is found instead of the conserved basic residue two positions before the second zinc-coordinating histidine on the ZF 5 helix, and the linker sequence is DTDKE in place of the classical TGEKP sequence. In a canonical ßßα fold, such unusual primary structure elements should cause severe electrostatic repulsions. In this context, we have characterized the structure and the dynamics of a Miz-1 construct comprising ZFs 5-8 (Miz 5-8) by solution-state NMR. Whilst ZFs 5, 7 and 8 were shown to adopt the classical ßßα fold for C2H2 ZFs, the number of long-range NOEs was insufficient to define a classical fold for ZF 6. We show by using (15)N-relaxation dispersion experiments that this lack of NOEs is due to the presence of extensive motions on the µs-ms timescale. Since this negatively charged region would have to be located near the phosphodiester backbone in a DNA complex, we propose that in addition to promoting conformational searches, it could serve as a hinge region to keep ZFs 1-4 away from DNA.

Primary and Secondary Progressive Aphasia in Posterior Cortical Atrophy.

BACKGROUND: Posterior cortical atrophy (PCA) is a clinico-radiological syndrome characterized by a progressive decline in visuospatial/visuoperceptual processing. PCA is accompanied by the impairment of other cognitive functions, including language abilities. METHODS: The present study focused on three patients presenting with language complaints and a clinical profile that was compatible with PCA. In addition to neurological and neuroimaging examinations, they were assessed with comprehensive batteries of neuropsychological and neurolinguistic tests. RESULTS: The general medical profile of the three patients is consistent with PCA, although they presented with confounding factors, making diagnosis less clear. The cognitive profile of the three patients was marked by Balint and Gerstmann's syndromes as well as impairments affecting executive functions, short-term and working memory, visuospatial and visuoperceptual abilities, and sensorimotor execution abilities. Their language ability was characterized by word-finding difficulties and impairments of sentence comprehension, sentence repetition, verbal fluency, narrative speech, reading, and writing. CONCLUSIONS: This study confirmed that PCA is marked by visuospatial and visuoperceptual deficits and reported evidence of primary and secondary language impairments in the three patients. The similarities of some of their language impairments with those found in the logopenic variant of primary progressive aphasia is discussed from neurolinguistic and neuroanatomical points of view.

Influence of Ca2+ and pH on the folding of the prourotensin II precursor.

Proper folding is a crucial step for the trafficking of proteins through the secretory pathway. We hypothesized that the secretory granules of endocrine cells provide optimal folding conditions of prohormone precursors for cleavage. Here, using circular dichroism and in vitro processing on purified prourotensin II (ProUII), we show that the precursor undergoes pH- and Ca(2+)-dependent conformational and stability changes. ProUII has a stable tertiary structure at pH 5.5 in presence of Ca(2+) and is correctly cleaved in these conditions by prohormone convertases. Taken together, our results support the notion that precursors may need to be optimally folded in the lumen of secretory granules for their processing.