scholarly journals Intermediate Filament Protein Domain Interactions as Revealed by Two-hybrid Screens

1996 ◽  
Vol 271 (3) ◽  
pp. 1599-1604 ◽  
Author(s):  
Jin-jun Meng ◽  
Sohaib Khan ◽  
Wallace Ip
Keyword(s):  
Intermediate Filament ◽  
Protein Domain ◽  
Intermediate Filament Protein ◽  
Domain Interactions ◽  
Filament Protein ◽  
Two Hybrid

scholarly journals The Saccharomyces cerevisiae 14-3-3 protein Bmh2 is required for regulation of the phosphorylation status of Fin1, a novel intermediate filament protein

2002 ◽  
Vol 365 (1) ◽  
pp. 51-56 ◽  
Author(s):  
Isabel MAYORDOMO ◽  
Pascual SANZ
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Transcriptional Regulation ◽  
Intermediate Filament ◽  
Protein Phosphatase ◽  
Catalytic Subunit ◽  
Protein Phosphatase 1 ◽  
Intermediate Filament Protein ◽  
Filament Protein ◽  
Two Hybrid ◽  
Hybrid Screening

In order to identify proteins that interact with Bmh2, a yeast member of the 14-3-3 protein family, we performed a two-hybrid screening using LexA-Bmh2 as bait. We identified Fin1, a novel intermediate filament protein, as the protein that showed the highest degree of interaction. We also identified components of the vesicular transport machinery such as Gic2 and Msb3, proteins involved in transcriptional regulation such as Mbf1, Gcr2 and Reg2, and a variety of other different proteins (Ppt1, Lre1, Rps0A and Ylr177w). We studied the interaction between Bmh2 and Fin1 in more detail and found that Bmh2 only interacted with phosphorylated forms of Fin1. In addition, we showed that Glc7, the catalytic subunit of the protein phosphatase 1 complex, was also able to interact with Fin1.

scholarly journals The Intermediate Filament Protein Vimentin Is a New Target for Epigallocatechin Gallate

2005 ◽  
Vol 280 (17) ◽  
pp. 16882-16890 ◽  
Author(s):  
Svetlana Ermakova ◽  
Bu Young Choi ◽  
Hong Seok Choi ◽  
Bong Seok Kang ◽  
Ann M. Bode ◽  
...  
Keyword(s):  
Intermediate Filament ◽  
Epigallocatechin Gallate ◽  
Intermediate Filament Protein ◽  
Filament Protein

A cDNA from Drosophila melanogaster encodes a lamin C-like intermediate filament protein

1993 ◽  
Vol 104 (4) ◽  
pp. 1263-1272 ◽  
Author(s):  
C.A. Bossie ◽  
M.M. Sanders
Keyword(s):  
Drosophila Melanogaster ◽  
Intermediate Filament ◽  
Blot Analysis ◽  
Polytene Chromosomes ◽  
Intermediate Filament Protein ◽  
Chromosome 2 ◽  
Cross Hybridization ◽  
Intermediate Filament Proteins ◽  
Nuclear Lamins ◽  
Filament Protein

A novel intermediate filament cDNA, pG-IF, has been isolated from a Drosophila melanogaster embryonic expression library screened with a polyclonal antiserum produced against a 46 kDa cytoskeletal protein isolated from Kc cells. This 46 kDa protein is known to be immunologically related to vertebrate intermediate filament proteins. The screen resulted in the isolation of four different cDNA groups. Of these, one has been identified as the previously characterized Drosophila nuclear lamin cDNA, Dm0, and a second, pG-IF, demonstrates homology to Dm0 by cross hybridization on Southern blots. DNA sequence analysis reveals that pG-IF encodes a newly identified intermediate filament protein in Drosophila. Its nucleotide sequence is highly homologous to nuclear lamins with lower homology to cytoplasmic intermediate filament proteins. pG-IF predicts a protein of 621 amino acids with a predicted molecular mass of 69,855 daltons. In vitro transcription and translation of pG-IF yielded a protein with a SDS-PAGE estimated molecular weight of approximately 70 kDa. It contains sequence principles characteristic of class V intermediate filament proteins. Its near neutral pI (6.83) and the lack of a terminal CaaX motif suggests that it may represent a lamin C subtype in Drosophila. In situ hybridization to polytene chromosomes detects one band of hybridization on the right arm of chromosome 2 at or near 51A. This in conjunction with Southern blot analysis of various genomic digests suggests one or more closely placed genes while Northern blot analysis detects two messages in Kc cells.

The presence or absence of a vimentin-type intermediate filament network affects the shape of the nucleus in human SW-13 cells

1994 ◽  
Vol 107 (6) ◽  
pp. 1593-1607 ◽  
Author(s):  
A.J. Sarria ◽  
J.G. Lieber ◽  
S.K. Nordeen ◽  
R.M. Evans
Keyword(s):  
Electron Microscopy ◽  
Intermediate Filament ◽  
Nuclear Morphology ◽  
Intermediate Filament Protein ◽  
Mosaic Pattern ◽  
Expression Plasmid ◽  
Vimentin Expression ◽  
Filament System ◽  
Filament Protein ◽  
Filament Network

Human SW-13 cells express the intermediate filament protein vimentin in a mosaic pattern (Hedberg, K. K. and Chen, L. B. (1986). Exp. Cell Res. 163, 509–517). We have isolated SW-13 clones that do (vim+) or do not (vim-) synthesize vimentin as analyzed using anti-intermediate filament immunofluorescence, electron microscopy and two-dimensional gel analysis of detergent-extracted preparations. Vimentin is the only cytoplasmic intermediate filament protein present in the vim+ cells, and the vim- cells do not contain any detectable cytoplasmic intermediate filament system. The presence or absence of intermediate filaments did not observably affect the distribution of mitochondria, endoplasmic reticulum, microtubules or actin stress fibers when these structures were visualized by fluorescence microscopy. However, electron microscopy and anti-lamin A/C immunofluorescence studies showed that nuclear morphology in vim- cells was frequently characterized by large folds or invaginations, while vim+ cells had a more regular or smooth nuclear shape. When vim- cells were transfected with a mouse vimentin expression plasmid, the synthesis of a mouse vimentin filament network restored the smooth nuclear morphology characteristic of vim+ cells. Conversely, when vim+ cells were transfected with a carboxy-terminally truncated mutant vimentin, expression of the mutant protein disrupted the organization of the endogenous vimentin filaments and resulted in nuclei with a prominently invaginated morphology. These results indicated that in SW-13 cells the vimentin filament system affects the shape of the nucleus.

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