Search

The 26S proteasome, composed of the 20S core and the 19S regulatory complex, plays a central role in ubiquitin-dependent proteolysis by catalyzing degradation of polyubiquitinated proteins. In a search for proteins involved in regulation of the proteasome, we affinity purified the 19S regulatory complex from HeLa cells and identified a novel protein of 43 kDa in size as an associated protein. Immunoprecipitation analyses suggested that this protein specifically interacted with the proteasomal ATPases. Hence the protein was named proteasomal ATPase-associated factor 1 (PAAF1). Immunoaffinity purification of PAAF1 confirmed its interaction with the 19S regulatory complex and further showed that the 19S regulatory complex bound with PAAF1 was not stably associated with the 20S core. Overexpression of PAAF1 in HeLa cells decreased the level of the 20S core associated with the 19S complex in a dose-dependent fashion, suggesting that PAAF1 binding to proteasomal ATPases inhibited the assembly of the 26S proteasome. Proteasomal degradation assays using reporters based on green fluorescent protein revealed that overexpression of PAAF1 inhibited the proteasome activity in vivo. Furthermore, the suppression of PAAF1 expression that is mediated by small inhibitory RNA enhanced the proteasome activity. These results suggest that PAAF1 functions as a negative regulator of the proteasome by controlling the assembly/disassembly of the proteasome. [ABSTRACT FROM AUTHOR]

Studies proteins involved in the regulation of cullin-containing E3 ubiquitin ligases by immunopurifying CUL4B-containing complex from HeLa cells and by identifying TIP120A as an associated protein by mass spectrometry. Role of cullin-containing ubiquitin-protein isopeptide ligases in regulating the abundance of key proteins involved in cellular processes such as cell cycle and cytokine signaling; Stability of the interaction of TIP120A with cullin family proteins.

Internal auditory canal (IAC) metastasis due to leptomeningeal carcinomatosis (LMC) from gastric cancer (GC) has rarely been reported. Early manifestation of symptoms, such as hearing loss, vertigo and facial paralysis, in cases of IAC metastasis due to LMC may facilitate the early detection of brain metastasis. To the best of our knowledge, the present study is the first to report IAC metastasis due to LMC in human epidermal growth factor receptor 2 (Her2)-positive GC. This study reports a case of an Her2-positive GC patient with LMC including IAC metastasis, who presented with acute sensorineural hearing loss, ipsilateral facial paralysis and vertigo during trastuzumab containing chemotherapy. The current study also discusses the early diagnosis and management of this complicated condition, demonstrating that clinical suspicion is key for a prompt diagnosis and proper management of LMC including IAC metastasis in Her2-positive GC. [ABSTRACT FROM AUTHOR]