A monoclonal antibody which recognises each of the nuclear lamin polypeptides in mammalian cells.

Burke, Brian; Tooze, John; Warren, Graham

doi:10.1002/j.1460-2075.1983.tb01431.x

Cited by 92 publications

(33 citation statements)

References 24 publications

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“…Conventional fluorescence and confocal laser scanning microscopy was used to confirm antibody localization of lamin B in ethanol-fixed cells. Lamin B staining was confined to the nuclear periphery (not shown), as has been observed by others (4,6), and confirmed our ability to preserve the antigenicity and three-dimensional localization of NMPs during sample preparation.…”

Section: Optimization Of Staining Proceduressupporting

confidence: 89%

Flow cytometric analysis of nuclear matrix proteins: Method and potential applications

et al. 1996

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Section: Optimization Of Staining Proceduressupporting

confidence: 89%

Flow cytometric analysis of nuclear matrix proteins: Method and potential applications

et al. 1996

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“…Primary antibodies used were as follows: rabbit polyclonal to rab6 C-19 peptide (Santa Cruz Biotechnology, Santa Cruz, CA); sheep polyclonal to human TGN46 (Serotec, Oxford, United Kingdom); mouse anti-TGN38 (Affinity Bioreagents, Golden, CO); mouse monoclonal against human transferrin receptor (Zymed Laboratories, South San Francisco, CA); mouse monoclonal ␥-adaptin (clone 100.3; Sigma-Aldrich); mouse monoclonal to early endosomal antigen-1 (EEA)-1 (BD Transduction Laboratories, Lexington, KY); lysosomal associated membrane protein-1 (LAMP1) (H4A3) monoclonal antibody (mAb) (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA); mouse monoclonal 9E10 recognizing the myc tag epitope (Covance, Berkeley, CA) and mouse anti-␣-tubulin (DM1A; Sigma-Aldrich). Noncommercial antibodies used were rabbit anti-p24␥ 3 (gp27; Fullekrug et al, 1999), polyclonal anti-Sar1p antibody (Pepperkok laboratory; unpublished data), polyclonal anti-GFP (Karsenti laboratory; unpublished data) and from the following sources (given in parentheses): rabbit antisera to the cytoplasmic domain of human 46-kDa cation-dependent mannose 6-phosphate receptor (Annette Hille-Rehfeld, Gottingen University, Germany; Klumperman et al, 1993); mouse monoclonal 53FC3 against mannosidase II (Graham Warren, Yale University, New Haven, CT; Burke et al, 1983), rabbit anti-rat mannosidase II (Kelly Moremen, Georgia University, Athens, GA; Moremen and Touster, 1985), monoclonal anti-GalT (Ulla Mandel, Copenhagen University, Copenhagen, Denmark; Almeida et al, 1999), sheep anti-GM130 (Francis Barr, MPI-Martinsried, Germany; published data), sheep antigolgin-84 (Martin Lowe, Manchester University, Manchester, United Kingdom; Diao et al, 2003), rabbit anti-sec61␤ (Bernard Dobberstein, Heidelberg University, Heidelberg, Germany; High et al, 1993), monoclonal 5B10 anti-rab6 (Angelika Barnekow, Muenster University, Muenster, Germany; Schiedel et al, 1995), monoclonal 50.1 anti-dynamitin (Richard Vallee, Columbia University, New York, NY; Paschal et al, 1993), and rabbit polyclonal anti-KAP3 (Tobias Stauber, European Molecular Biology Laboratory, Heidelberg, Germany; unpublished data). Secondary antibodies to sheep, rabbit, and mouse conjugated to were obtained from Molecular Probes (Eugene, OR).…”

Section: Materials and Antibodiesmentioning

confidence: 99%

Regulation of Microtubule-dependent Recycling at theTrans-Golgi Network by Rab6A and Rab6A'

Young

Stauber

Nery

et al. 2005

MBoC

104

126

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The small GTPase rab6A but not the isoform rab6A has previously been identified as a regulator of the COPIindependent recycling route that carries Golgi-resident proteins and certain toxins from the Golgi to the endoplasmic reticulum (ER). The isoform rab6A has been implicated in Golgi-to-endosomal recycling. Because rab6A but not A , binds rabkinesin6, this motor protein is proposed to mediate COPI-independent recycling. We show here that both rab6A and rab6A GTP-restricted mutants promote, with similar efficiency, a microtubule-dependent recycling of Golgi resident glycosylation enzymes upon overexpression. Moreover, we used small interfering RNA mediated down-regulation of rab6A and A' expression and found that reduced levels of rab6 perturbs organization of the Golgi apparatus and delays Golgi-to-ER recycling. Rab6-directed Golgi-to-ER recycling seems to require functional dynactin, as overexpression of p50/dynamitin, or a C-terminal fragment of Bicaudal-D, both known to interact with dynactin inhibit recycling. We further present evidence that rab6-mediated recycling seems to be initiated from the trans-Golgi network. Together, this suggests that a recycling pathway operates at the level of the trans-Golgi linking directly to the ER. This pathway would be the preferred route for both toxins and resident Golgi proteins. INTRODUCTIONAnterograde flow of material through the exocytic pathway is offset by the constant recycling of both proteins and lipids. This recycling helps to ensure that steady-state distributions of resident proteins are maintained within the pathway at the same time as keeping a lipid balance. Besides intra-Golgi recycling between cisternae (Hoe et al., 1995;Love et al., 1998;Lanoix et al., 1999;Lin et al., 1999), Golgi glycosylation enzymes also can recycle directly to the endoplasmic reticulum (ER). It is estimated that at least 5% of Golgi resident enzymes reside in the ER, at steady state (Pelletier et al., 2000). This pool constitutes recycled material originating from the Golgi apparatus (Cole et al., 1998;Storrie et al., 1998).The extent of ER recycling of Golgi resident membrane proteins is sufficient to allow for complete assembly of functional Golgi stacks. Thus, upon addition of nocodazole to depolymerize microtubules, the central and juxtanuclear Golgi apparatus relocates to the 100 or so peripheral ER exit sites scattered throughout the cell. This microtubule-independent relocation occurs in the absence of any detectable elements tracking outwards from the central Golgi apparatus. Rather, the Golgi apparatus undergoes a molecular deconstruction in the trans-to-cis-direction, i.e., resident glycosylation enzymes of the trans-cisternae/trans-Golgi network (TGN) relocate with faster kinetics than do enzymes of the medial-cisternae (Yang and Storrie, 1998). Enzymes seemingly enter the ER close to the microtubule organizing center (MTOC), diffuse through the ER, and then reemerge at peripheral ER exit sites. Here, Golgi stacks are then reassembled into discrete, yet fully functiona...

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Section: Introductionmentioning

confidence: 99%

“…Peptide maps of lamins A and C are similar, but they are different from those of lamin B (6). However, certain monoclonal antibodies have been shown to crossreact with all three lamins, indicating that some epitopes are shared by all three lamins (7,8).The lamins have been shown to be synthesized throughout the cell cycle (9). In vitro translation of mRNA (10) showed that lamins A and C are synthesized from separate mRNAs with lamin A being made as a precursor approximately 2 kDa larger than mature lamin A.…”

mentioning

confidence: 99%

cDNA sequencing of nuclear lamins A and C reveals primary and secondary structural homology to intermediate filament proteins.

Fisher¹,

Chaudhary²,

Blobel³

1986

Proc. Natl. Acad. Sci. U.S.A.

637

405

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The amino acid sequences. deduced from cDNA clones of human lamin A and lamin C show identity between these two lamins except for an extra 9.0-kDa carboxylterminal tail that is present only in lamin A. Both latins A and C contain an a-helical domain of approximately 360 residues that shows striking homology to a corresponding a-helical rod domain that, is the structural hallmark of all intermediate filament proteins. How'ever, the lamin a-helical domain Implications of the presence of these and other domains in lamins A and C for the assembly of the nuclear lamina are discussed.The nuclear lamina is a polymeric structure that is intercalated between chromatin and the inner membrane of the nuclear envelope. In mammalian cells the lamina consists primarily ofthree proteins (1-4) that have been termed lamins A (70 kDa), B (67 kDa), and C (60 kDa) (5). Peptide maps of lamins A and C are similar, but they are different from those of lamin B (6). However, certain monoclonal antibodies have been shown to crossreact with all three lamins, indicating that some epitopes are shared by all three lamins (7,8).The lamins have been shown to be synthesized throughout the cell cycle (9). In vitro translation of mRNA (10) showed that lamins A and C are synthesized from separate mRNAs with lamin A being made as a precursor approximately 2 kDa larger than mature lamin A. The precursor is converted into mature lamin A only after incorporation into the lamina structure, as shown by pulse-chase experiments (9).The lamina is depolymerized during mitotic prophase by a process that may involve transient hyperphosphorylation (5).Concomitant with a gradual dephosphorylation, the lamins repolymerize in telophase (3,5). How the regular protein meshwork (2) that constitutes the lamina is assembled from the three lamins is not known. The lamins must contain domains that are involved in this polymerization and that interact with chromatin on the nucleoplasmic face of the lamina, with the inner nuclear membrane on the other face of the lamina, and, most likely, with nuclear pore complexes (1, 2). As such domains could be revealed from the amino acid sequences of the lamins, we decided to clone their cDNAs. We report here the deduced amino acid sequences for lamins A and C. While The publication costs of this article were defrayed in part by page charge payment., This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

show abstract

A monoclonal antibody which recognises each of the nuclear lamin polypeptides in mammalian cells.

Cited by 92 publications

References 24 publications

Flow cytometric analysis of nuclear matrix proteins: Method and potential applications

Flow cytometric analysis of nuclear matrix proteins: Method and potential applications

Regulation of Microtubule-dependent Recycling at theTrans-Golgi Network by Rab6A and Rab6A'

cDNA sequencing of nuclear lamins A and C reveals primary and secondary structural homology to intermediate filament proteins.

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