Retention and mobility of the mammalian lamin B receptor in the plant nuclear envelope.
BACKGROUND
In a previous study, we showed that GFP (green fluorescent protein) fused to the N-terminal 238 amino acids of the mammalian LBR (lamin B receptor) localized to the NE (nuclear envelope) when expressed in the plant Nicotiana tabacum. The protein was located in the NE during interphase and migrated with nuclear membranes during cell division. Targeting and retention of inner NE proteins requires several mechanisms: signals that direct movement through the nuclear pore complex, presence of a transmembrane domain or domains and retention by interaction with nuclear or nuclear-membrane constituents.


RESULTS
Binding mutants of LBR-GFP were produced to investigate the mechanisms for the retention of LBR in the NE. FRAP (fluorescence recovery after photobleaching) analysis of mutant and wild-type constructs was employed to examine the retention of LBR-GFP in the plant NE. wtLBR-GFP (wild-type LBR-GFP) was shown to have significantly lower mobility in the NE than the lamin-binding domain deletion mutant, which showed increased mobility in the NE and was also localized to the endoplasmic reticulum and punctate structures in some cells. Modification of the chromatin-binding domain resulted in the localization of the protein in nuclear inclusions, in which it was immobile.


CONCLUSIONS
As expression of truncated LBR-GFP in plant cells results in altered targeting and retention compared with wtLBR-GFP, we conclude that plant cells can recognize the INE (inner NE)-targeting motif of LBR. The altered mobility of the truncated protein suggests that not only do plant cells recognize this signal, but also have nuclear proteins that interact weakly with LBR.
Graumann
Katja
K
Research School of Life Sciences, Oxford Brookes University, Gipsy Lane, Oxford OX3 0BP, UK.
Irons
Sarah L
SL
Runions
John
J
Evans
David E
DE
D000818:Animals; D037281:Calnexin / Q000235:genetics / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D036681:Fluorescence Recovery After Photobleaching; D006801:Humans; D045586:Intranuclear Inclusion Bodies / Q000378:metabolism / Q000648:ultrastructure; D009685:Nuclear Envelope / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology*; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana / Q000166:cytology* / Q000378:metabolism; D000097682:Lamin B Receptor
D037281:Calnexin; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins; D000097682:Lamin B Receptor