Nuclear envelope transmembrane proteins (NETs) that are up-regulated during myogenesis.
BACKGROUND
The nuclear lamina is a protein meshwork lining the inner nuclear membrane, which contains a polymer of nuclear lamins associated with transmembrane proteins of the inner nuclear membrane. The lamina is involved in nuclear structure, gene expression, and association of the cytoplasmic cytoskeleton with the nucleus. We previously identified a group of 67 novel putative nuclear envelope transmembrane proteins (NETs) in a large-scale proteomics analysis. Because mutations in lamina proteins have been linked to several human diseases affecting skeletal muscle, we examined NET expression during differentiation of C2C12 myoblasts. Our goal was to identify new nuclear envelope and lamina components whose expression is coordinated with muscle differentiation.


RESULTS
Using transcriptional microarray analysis, we found that expression of 6 of the NETs significantly increases during myoblast differentiation. We confirmed these results using quantitative RT-PCR, and furthermore, found that all 6 NETs are expressed at high levels in adult mouse skeletal muscle relative to 9 other tissues examined. Using epitope-tagged cDNAs, we determined that the 5 NETs we could analyze (NETs 9, 25, 32, 37 and 39) all target to the nuclear envelope in C2C12 cells. Furthermore, the 3 NETs that we could analyze by immunoblotting were highly enriched in nuclear envelopes relative to microsomal membranes purified from mouse liver. Database searches showed that 4 of the 6 up-regulated NETs contain regions of homology to proteins previously linked to signaling.


CONCLUSIONS
This work identified 6 NETs that are predicted to have important functions in muscle development and/or maintenance from their expression patterns during myoblast differentiation and in mouse tissues. We confirmed that 5 of these NETs are authentic nuclear envelope proteins. Four members of this group have potential signaling functions at the NE, based on their sequence homologies.
Chen
I-Hsiung Brandon
IH
Department of Cell Biology, The Scripps Research Institute, La Jolla CA 92037, USA. ibchen@yahoo.com <ibchen@yahoo.com>
Huber
Michael
M
Guan
Tinglu
T
Bubeck
Anja
A
Gerace
Larry
L
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D008099:Liver / Q000166:cytology; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics*; D051379:Mice; D008958:Models, Molecular; D024510:Muscle Development / Q000235:genetics*; D009124:Muscle Proteins / Q000737:chemistry / Q000235:genetics*; D018482:Muscle, Skeletal / Q000378:metabolism; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism*; D009685:Nuclear Envelope / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D015854:Up-Regulation
D008565:Membrane Proteins; D009124:Muscle Proteins; D009687:Nuclear Proteins