Target Acquisition in Multiscale Electronic Worlds

Since the advent of graphical user interfaces, electronic information has grown exponentially, whereas the size of screen displays has stayed almost the same. Multiscale interfaces were designed to address this mismatch, allowing users to adjust the scale at which they interact with information objects. Although the technology has progressed quickly, the theory has lagged behind. Multiscale interfaces pose a stimulating theoretical challenge, reformulating the classic target-acquisition problem from the physical world into an infinitely rescalable electronic world. We address this challenge by extending Fitts’ original pointing paradigm: we introduce the scale variable, thus defining a multiscale pointing paradigm. This article reports on our theoretical and empirical results. We show that target-acquisition performance in a zooming interface must obey Fitts’ law, and more specifically, that target-acquisition time must be proportional to the index of difficulty. Moreover, we complement Fitts’ law by accounting for the effect of view size on pointing performance, showing that performance bandwidth is proportional to view size, up to a ceiling effect. The first empirical study shows that Fitts’ law does apply to a zoomable interface for indices of difficulty up to and beyond 30 bits, whereas classical Fitts’ law studies have been confined in the 2-10 bit range. The second study demonstrates a strong interaction between view size and task difficulty for multiscale pointing, and shows a surprisingly low ceiling. We conclude with implications of these findings for the design of multiscale user interfaces

Targeting of cytochrome b2 into the mitochondrial intermembrane space

Cytochrome b2 contains 2-fold targeting information: an amino-terminal signal for targeting to the mitochondrial matrix, followed by a second cleavable sorting signal that functions in directing the precursor into the mitochondrial intermembrane space. The role of the second sorting sequence was analyzed by replacing one, two or all of the three positively charged amino acid residues which are present at the amino-terminal side of the hydrophobic core by uncharged residues or an acidic residue. With a number of these mutant precursor proteins, processing to the mature form was reduced or completely abolished and at the same time targeting to the matrix space occurred. The accumulation in the matrix depended on a high level of intramitochondrial ATP. At low levels of matrix ATP, the mutant proteins were sorted into the intermembrane space like the wild-type precursors. The results: (i) suggest the existence of one or more matrix components that specifically recognize the second sorting signal and thereby trigger the translocation into the intermembrane space; (ii) indicate that the mutant signals have reduced ability to interact with the recognition component(s) and then embark on the default pathway into the matrix by interacting with mitochondrial hsp70 in conjunction with matrix ATP; (iii) strongly argue against a mechanism by which the hydrophobic segment of the sorting sequence stops translocation in the hydrophobic phase of the inner membrane

Characterization of the mitochondrial processing peptidase of Neurospora crassa

The mitochondrial processing peptidase (MPP) of Neurospora crassa is constituted by an alpha- and a beta-subunit. We have purified alpha-MPP after expression in Escherichia coli while beta-MPP was purified from mitochondria. A fusion protein between precytochrome b2 and mouse dihydrofolate reductase was expressed in E. coli, and the purified protein was used as substrate for MPP. Both subunits of MPP are required for processing. MPP removes the matrix targeting signal of cytochrome b2 by a single cut, and the resulting presequence peptide is 31 amino acid residues in length. It acts as a competitive inhibitor of processing but has a approximately 30-fold lower affinity for MPP than the preprotein. Competition assays show that MPP recognizes the COOH- terminal portion of the presequence of cytochrome b2 rather than the NH2-terminal part which has the potential to form an amphiphilic helix. Substitution of arginine in position -2 of the matrix targeting sequence of cytochrome b2 prevents processing but not import of a chimeric precursor. Substitution of the tyrosyl residue in position +1 also prevents processing, indicating that MPP interacts with sequences COOH-terminal to the cleavage site. Non-cleavable preprotein is still recognized by MPP. Our data suggest that processing peptidase and import machinery recognize distinct structural elements in preproteins which, however, can be overlapping

Translocation arrest by reversible folding of a precursor protein imported into mitochondria

Passage of precursor proteins through translocation contact sites of mitochondria was investigated by studying the import of a fusion protein consisting of the NH2-terminal 167 amino acids of yeast cytochrome b2 precursor and the complete mouse dihydrofolate reductase. Isolated mitochondria of Neurospora crassa readily imported the fusion protein. In the presence of methotrexate import was halted and a stable intermediate spanning both mitochondrial membranes at translocation contact sites accumulated. The complete dihydrofolate reductase moiety in this intermediate was external to the outer membrane, and the 136 amino acid residues of the cytochrome b2 moiety remaining after cleavage by the matrix processing peptidase spanned both outer and inner membranes. Removal of methotrexate led to import of the intermediate retained at the contact site into the matrix. Thus unfolding at the surface of the outer mitochondrial membrane is a prerequisite for passage through translocation contact sites. The membrane-spanning intermediate was used to estimate the number of translocation sites. Saturation was reached at 70 pmol intermediate per milligram of mitochondrial protein. This amount of translocation intermediates was calculated to occupy approximately 1% of the total surface of the outer membrane. The morphometrically determined area of close contact between outer and inner membranes corresponded to approximately 7% of the total outer membrane surface. Accumulation of the intermediate inhibited the import of other precursor proteins suggesting that different precursor proteins are using common translocation contact sites. We conclude that the machinery for protein translocation into mitochondria is present at contact sites in limited number

Espace et échelle du mouvement : la loi de Fitts dans le monde physique et dans les mondes électroniques

Nous avons conçu et mis en oeuvre une approche nouvelle du paradigme classique du pointage faisant jouer un rôle central à la variable d’échelle. Nous utilisons le diagramme espace-échelle introduit par Furnas et Bederson (1995) dans le champ de l’interaction homme-machine (IHM). Fondé sur une géométrie inédite, ce type de représentation permet de visualiser les variations d’échelle en même temps que les déplacements dans l’espace, ce qui nous a permis d’analyser la cinématique de la navigation au sein des mondes électroniques multi-échelle (ou zoomables). Les travaux rapportés dans ce document se développent autour de la frontière qui sépare le monde réel et les mondes d’information. Le monde réel dans lequel s’inscrivent normalement nos mouvements est typiquement dynamique : il met en jeu la masse, la longueur et le temps. En revanche, les mondes d’information auxquels les humains sont exposés depuis à peine deux décennies sont d’essence purement cinématique, en ce sens qu’ils ignorent typiquement la masse, et donc les contraintes de force, d’énergie et de puissance. C’est la nature cinématique des mondes d’information qui explique que leurs contenus soient indéfiniment rescalables, propriété dont les objets du monde physique ne jouissent que dans une étroite mesure, comme on le sait depuis Galilée

Central serotonin-2A (5-HT2A) receptor dysfunction in depression and epilepsy : the missing link?

5-Hydroxytryptamine 2A receptors (5-HT2A-Rs) are G-protein coupled receptors. In agreement with their location in the brain, they have been implicated not only in various central physiological functions including memory, sleep, nociception, eating and reward behaviors, but also in many neuropsychiatric disorders. Interestingly, a bidirectional link between depression and epilepsy is suspected since patients with depression and especially suicide attempters have an increased seizure risk, while a significant percentage of epileptic patients suffer from depression. Such epidemiological data led us to hypothesize that both pathologies may share common anatomical and neurobiological alteration of the 5-HT2A signaling. After a brief presentation of the pharmacological properties of the 5-HT2A-Rs, this review illustrates how these receptors may directly or indirectly control neuronal excitability in most networks involved in depression and epilepsy through interactions with the monoaminergic, GABAergic and glutamatergic neurotransmissions. It also synthetizes the preclinical and clinical evidence demonstrating the role of these receptors in antidepressant and antiepileptic responses.peer-reviewe

Protein folding causes an arrest of preprotein translocation into mitochondria in vivo

With vital yeast cells, a hybrid protein consisting of the amino- terminal third of the precursor to cytochrome b2 and of the entire dihydrofolate reductase was arrested on the import pathway into mitochondria. Accumulation of the protein in the mitochondrial membranes was achieved by inducing a stable tertiary structure of the dihydrofolate reductase domain. Thereby, three salient features of mitochondrial protein uptake in vivo were demonstrated: its posttranslational character; the requirement for unfolding of precursors; and import through translocation contact sites. The permanent occupation of translocation sites by the fusion protein inhibited the import of other precursors; it did, however, not lead to leakage of mitochondrial ions, implying the existence of a channel that is sealed around the membrane spanning polypeptide segment

Energy requirements for unfolding and membrane translocation of precursor proteins during import into mitochondria

ATP is involved in conferring transport competence to numerous mitochondrial precursor proteins in the cytosol. Unfolded precursor proteins were found not to require ATP for import into mitochondria, suggesting a role of ATP in the unfolding of precursors. Here we report the unexpected finding that a hybrid protein containing the tightly folded passenger protein dihydrofolate reductase becomes unfolded and specifically translocated across the mitochondrial membranes independently of added ATP. Moreover, interaction of the precursor with the mitochondrial receptor components does not require ATP. The results suggest that ATP is not involved in the actual process of unfolding during membrane translocation of precursors. ATP rather appears to be necessary for preventing the formation of improper structures of precursors in the cytosol and for folding of imported polypeptides on (and release from) chaperone-like molecules in the mitochondrial matrix