The p53-induced factor Ei24 inhibits nuclear import through an importin β–binding–like domain

Abstract

The p53-induced protein Ei24 can bind specifically to importin-β1 and importin-α2 to impede their normal role in nuclear import.

The ataxin-1 interactome reveals direct connection with multiple disrupted nuclear transport pathways

“…We set out to evaluate the effect of short-term expression of polyQ-ataxin-1, initially by assessing NPC distribution using the anti-nucleoporin (Nup) Mab414 antibody; no marked changes in staining were observed upon polyQ-ataxin-1 expression ( Figure 6A) indicating no wide-spread disruption of NPC distribution in the nuclear envelope. Indicative of general NPC functionality, nuclear accumulation of the transcription factor AF10, known to be importin/Ran-independent and to occur via direct interaction with NPC components 44,45 was not affected by polyQ-ataxin-1 ( Figure S4). Nucleoporin protein NUP62, within the central core of the NPC 46, 47 , was not altered in the presence of polyQ-ataxin-1 either in the presence or absence of arsenite stress ( Figure S5), in contrast to the impact of mutant huntingtin in HD 16 .…”

ULK2 Ser 1027 Phosphorylation by PKA Regulates Its Nuclear Localization Occurring through Karyopherin Beta 2 Recognition of a PY-NLS Motif

“…Fig 1D emphasizes the co-localization of ULK2 and Kapβ2 (white spot) in the enlarged image of the specific merged region. Pearson's correlation coefficient (PCC) of the co-localization between ULK2 and Kapβ2 was determined using an LSM710 [ 17 , 18 ]. The PCC value (0.60+/− 0.12; n = 5) indicates that the coexistence of ULK2 and Kapβ2 in HEK293 cells positively occurs with roughly a 60% probability.…”

“…Using Profile in the ZEN program which was provided by the manufacturer, the co-localizations of proteins were observed and confocal microscopic images scanned. The nuclear or cytoplasmic fluorescence intensity profile of ULK1/2 was evaluated from the fluorescence images, and the nuclear-to-total cell fluorescence (Fn/t) ratio was obtained using the ZEN program [ 13 , 17 , 18 ]. Pearson's correlation coefficient (PCC) of the co-localization between ULK2 and Kapβ2 was measured with an LSM710 (Zeiss, Germany).…”

Inhibition of ULK2 Autophagic Activity by Its Association with YAP

“…The co-localization analysis was performed on digitized CLSM images using ZEN2012 software (ZEISS, Germany), where a defined ULK2 threshold signal above the background level (secondary antibody alone control) was overlaid onto that of the IMP signal to produce a merged image to colocalized the ULK2 and YAP pixels. The pixel intensity of the merged image was expressed as a percentage of the total YAP signal to calculate the mean percentage of YAP colocalized with ULK2 (Barlow, Macleod, Noppen, Sanderson, & Guerin, 2010;Lieu et al, 2014). Pearson's correlation coefficient (PCC) of the co-localization between Yap and ULK2 was measured with an LSM710 microscope.…”

“…WIPI also mainly co-localized with the mutant protein in the cytoplasm in cells transfected with ULK2 PA ( Figure 4B), compared with WIPI in ULK2 WT-( Figure 4A) or P242A-mutant ( Figure 4C) transfected cells. PCC of the co-localization between ULK2 and WIPI was provided on the right side (Barlow et al, 2010;Lieu et al, 2014). PCC between the PA mutant and WIPI was 0.90 ± 0.04; n = 10), which was twice than that of ULK WT (0.56 ± 0.08; n = 10) or the P242A mutant (0.59 ± 0.05; n = 10).…”