Colocalization of intranuclear lamin foci with RNA splicing factors.
The lamins form a fibrous network underlying the inner nuclear membrane termed the nuclear lamina. In order to gain insights into the role of lamins in nuclear organization, we have characterized a monoclonal antibody (LA-2H10) raised against recombinant rat lamin A that labels nuclei in a speckled pattern in all cells of unsynchronized populations of HeLa and rat F-111 fibroblast cells, unlike the typical nuclear periphery staining by another monoclonal antibody to lamin A, LA-2B3. In immunolocalization studies the lamin A speckles or foci were found to colocalize with the RNA splicing factors SC-35 and U5-116 kD, but not with p80 coilin found in coiled bodies. Lamin B1 was also associated with these foci. These foci dispersed when cells entered mitosis and reformed during anaphase. The differential reactivity of LA-2H10 and LA-2B3 was retained after nuclei were extracted with detergents, nucleases and salt to disrupt interactions of lamins with chromatin and other nuclear proteins. Using deletion fragments of recombinant lamin A, the epitope recognized by LA-2H10 was located between amino acids 171 and 246. Our findings are consistent with a structural role for lamins in supporting nuclear compartments containing proteins involved in RNA splicing.
Jagatheesan
G
G
Centre for Cellular and Molecular Biology, Hyderabad 500 007, India.
Thanumalayan
S
S
Muralikrishna
B
B
Rangaraj
N
N
Karande
A A
AA
Parnaik
V K
VK
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D002467:Cell Nucleus / Q000378:metabolism*; D018604:Epitope Mapping; D006367:HeLa Cells; D006801:Humans; D034904:Lamin Type A; D034921:Lamin Type B; D034882:Lamins; D008856:Microscopy, Fluorescence; D009687:Nuclear Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D012326:RNA Splicing; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017384:Sequence Deletion
D000911:Antibodies, Monoclonal; D034904:Lamin Type A; D034921:Lamin Type B; D034882:Lamins; D009687:Nuclear Proteins; D011994:Recombinant Proteins