Interference with the cytoplasmic tail of gp210 disrupts “close apposition” of nuclear membranes and blocks nuclear pore dilation

Drummond, Sheona P; Wilson, Katherine L.

doi:10.1083/jcb.200108145

Cited by 55 publications

(41 citation statements)

References 58 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…They are accompanied by a local distribution of proteins concentrated near the pore opening which appear to facilitate pore formation (Drummond and Wilson 2002). To describe an isolated nuclear pore, we assign a hypothetical spontaneous curvature distribution having the Gaussian form (see Fig.…”

Section: Nuclear Poresmentioning

confidence: 99%

Modeling protein-mediated morphology in biomembranes

Agrawal

Steigmann

2008

Biomech Model Mechanobiol

114

View full text Add to dashboard Cite

The equilibrium theory for lipid membranes is used to describe the structure of nuclear pores and the membrane shapes accompanying endocytosis. The commonly used variant of the theory contains a fixed parameter called the spontaneous curvature which accounts for asymmetry in the bending response of the membrane. This is replaced here by a variable distribution of spontaneous curvature representing the influence of attached proteins. The required adjustments to the standard theory are described and the resulting model is applied to the study of membrane morphology at the cites of protein-assisted nuclear pore formation and endocytosis.

show abstract

Section: Nuclear Poresmentioning

confidence: 99%

Modeling protein-mediated morphology in biomembranes

Agrawal

Steigmann

2008

Biomech Model Mechanobiol

114

View full text Add to dashboard Cite

show abstract

“…Three pore integral membrane proteins, POM121, NDC1, and gp210, exist in vertebrates. POM121 appears early in nuclear assembly, making its binding partners in the nuclear pore of particular interest (Gerace et al, 1982;Chaudhary and Courvalin, 1993;Hallberg et al, 1993;Bodoor et al, 1999;Drummond and Wilson, 2002;Antonin et al, 2005;Dultz et al, 2008). Furthermore, RNAi knockdown of POM121 has, in many but not all cases, shown POM121 to be required for nuclear pore formation (Antonin et al, 2005;Mansfeld et al, 2006;Funakoshi et al, 2007).…”

mentioning

confidence: 99%

Capture of AT-rich Chromatin by ELYS Recruits POM121 and NDC1 to Initiate Nuclear Pore Assembly

Rasala

Ramos

Harel

et al. 2008

MBoC

146

201

View full text Add to dashboard Cite

Assembly of the nuclear pore, gateway to the genome, from its component subunits is a complex process. In higher eukaryotes, nuclear pore assembly begins with the binding of ELYS/MEL-28 to chromatin and recruitment of the large critical Nup107-160 pore subunit. The choreography of steps that follow is largely speculative. Here, we set out to molecularly define early steps in nuclear pore assembly, beginning with chromatin binding. Point mutation analysis indicates that pore assembly is exquisitely sensitive to the change of only two amino acids in the AT-hook motif of ELYS. The dependence on AT-rich chromatin for ELYS binding is borne out by the use of two DNA-binding antibiotics. AT-binding Distamycin A largely blocks nuclear pore assembly, whereas GC-binding Chromomycin A 3 does not. Next, we find that recruitment of vesicles containing the key integral membrane pore proteins POM121 and NDC1 to the forming nucleus is dependent on chromatin-bound ELYS/Nup107-160 complex, whereas recruitment of gp210 vesicles is not. Indeed, we reveal an interaction between the cytoplasmic domain of POM121 and the Nup107-160 complex. Our data thus suggest an order for nuclear pore assembly of 1) AT-rich chromatin sites, 2) ELYS, 3) the Nup107-160 complex, and 4) POM121-and NDC1-containing membrane vesicles and/or sheets, followed by (5) assembly of the bulk of the remaining soluble pore subunits. INTRODUCTIONThe possession of a nuclear envelope (NE) that encompasses the genome is the defining characteristic of all eukaryotes. The envelope consists of double nuclear membranes, hundreds to thousands of nuclear pore complexes (NPCs), and in higher eukaryotes, a nuclear lamina. Bidirectional transport of protein and RNA molecules through the nuclear envelope is mediated exclusively by NPCs, large structures ϳ60 -125 MDa in size (Reichelt et al., 1990;Macara, 2001;Quimby and Corbett, 2001;Goldfarb et al., 2004;Pemberton and Paschal, 2005;Patel et al., 2007).In higher eukaryotes the nuclear envelope, including pore complexes, disassembles at mitosis as a prelude to spindle assembly and chromosome segregation (Burke and Ellenberg, 2002;Margalit et al., 2005;Prunuske et al., 2006). This disassembly then necessitates nuclear envelope reformation around each set of segregated chromosomes toward the end of mitosis, a process that involves both nuclear membrane recruitment and nuclear pore formation.Analysis of the pore subunits produced by mitotic disassembly has provided the most useful clues to nearest neighbor interactions within the vertebrate pore. Vertebrate nuclear pores are comprised of ϳ30 different proteins or nucleoporins (Nups) in 8-32 copies each, to give a 500-1000 protein structure (Cronshaw et al., 2002). At mitosis the massive vertebrate pore disassembles into ϳ14 soluble subunits, each with a distinct protein composition, whereas the integral membrane pore proteins, POM121, NDC1, and gp210, segregate into endoplasmic reticulum (ER) sheets and vesicles (Gerace et al., 1982;Wozniak et al., 1989;Greber et al., 1990;Hallberg ...

show abstract

“…The majority of gp210 is on the cisternal side of the NPC pore membrane, with only a short 60-aminoacid region exposed on the NPC side of the membrane (Greber et al 1990). Both overexpression of this tail domain and RNAi-mediated depletion of gp210 inhibit NPC assembly (Drummond and Wilson 2002;Cohen et al 2003). Expression of antibodies that associate with the lumenal domain of gp210 results in decreased nuclear import of NLS-containing proteins and a reduced diffusion rate across the pore of smaller proteins, implicating the glycosylated region of gp210 in NPC function (Greber and Gerace 1992).…”

mentioning

confidence: 99%

Nuclear Pore Complex Function in Saccharomyces cerevisiae Is Influenced by Glycosylation of the Transmembrane Nucleoporin Pom152p

et al. 2005

View full text Add to dashboard Cite

The regulated transport of proteins across the nuclear envelope occurs through nuclear pore complexes (NPCs), which are composed of .30 different protein subunits termed nucleoporins. While some nucleoporins are glycosylated, little about the role of glycosylation in NPC activity is understood. We have identified loss-of-function alleles of ALG12, encoding a mannosyltransferase, as suppressors of a temperature-sensitive mutation in the gene encoding the FXFG-nucleoporin NUP1. We observe that nup1D cells import nucleophilic proteins more efficiently when ALG12 is absent, suggesting that glycosylation may influence nuclear transport. Conditional nup1 and nup82 mutations are partially suppressed by the glycosylation inhibitor tunicamycin, while nic96 and nup116 alleles are hypersensitive to tunicamycin treatment, further implicating glycosylation in NPC function. Because Pom152p is a glycosylated, transmembrane nucleoporin, we examined genetic interactions between pom152 mutants and nup1D. A nup1 deletion is lethal in combination with pom152D, as well as with truncations of the Nterminal and transmembrane regions of Pom152p. However, truncations of the N-glycosylated, lumenal domain of Pom152p and pom152 mutants lacking N-linked glycosylation sites are viable in combination with nup1D, suppress nup1D temperature sensitivity, and partially suppress the nuclear protein import defects associated with the deletion of NUP1. These data provide compelling evidence for a role for glycosylation in influencing NPC function.

show abstract

Interference with the cytoplasmic tail of gp210 disrupts “close apposition” of nuclear membranes and blocks nuclear pore dilation

Cited by 55 publications

References 58 publications

Modeling protein-mediated morphology in biomembranes

Modeling protein-mediated morphology in biomembranes

Capture of AT-rich Chromatin by ELYS Recruits POM121 and NDC1 to Initiate Nuclear Pore Assembly

Nuclear Pore Complex Function in Saccharomyces cerevisiae Is Influenced by Glycosylation of the Transmembrane Nucleoporin Pom152p

Contact Info

Product

Resources

About