The Ras family of proteins and several of the nuclear lamins contain a carboxyl-terminal """"""""CaaX motif"""""""" that triggers protein isoprenylation and methylation. We have intensively studied these posttranslational modifications. We created gene-targeted mice to examine the role of all of the enzymes involved in the posttranslational processing of CaaX proteins [protein farnesyltransferase (FTase), protein geranylgeranyl- transferase-I (GGTase-I), ICMT, RCE1, ZMPSTE24]. We have examined the importance of each enzyme in the growth of Ras-induced tumors. We also discovered that ZMPSTE24 is a prelamin A protease and that its absence leads to an accumulation of farnesyl-prelamin A. This accumulation of farnesyl-prelamin A underlies certain progeroid disorders in humans, including Hutchinson-Gilford progeria syndrome. We created gene- targeted mice and investigated the importance of protein farnesylation in the pathogenesis and treatment of progeria. We also discovered that lamins B1 and B2 are crucial for neuronal migration in the brain. Our research program on CaaX proteins has been productive, resulting in 38 publications during the past 5 years. The objective of this renewal application is to continue our work on the in vivo importance of the modifications of CaaX proteins, focusing on specific topics relevant to human disease.
Specific Aim 1 is to use genetically modified mice to examine the importance of lamins B1 and B2 in the brain and in other tissues. Our preliminary studies suggest that B-type lamins are essential for neuronal migration. We need to investigate this issue in depth. Also, we need to determine whether the individual proteins, lamin B1 and lamin B2, have intrinsically different roles in this process. Finally, we need to investigate the roles of lamin B1 and lamin B2 in adult tissues. The fact that lamin B1 and lamin B2 knockout mice die at birth means that no one has yet explored the physiologic importance of these proteins in adult tissues. Very recently, we have created conditional knockout alleles for both Lmnb1 and Lmnb2, and over the next few years, we intend to use these tools to address the importance of the B-type lamins in adult tissues.
Specific Aim 2 is to define the importance of the posttranslational modifications of prelamin A in the assembly of the nuclear lamina, and to define the importance of protein farnesylation in the pathogenesis of progeroid syndromes. Over the past few years, our laboratory has generated knock-in mice that synthesize exclusively prelamin A and other knock-in mice that produce exclusively mature lamin A (where lamin A is produced directly, bypassing the posttranslational processing). Already, we have discovered differences in the assembly of the nuclear lamina in these mice. Further analyses of these mice should help us to define the physiologic importance of the posttranslational processing of prelamin A. Also, additional knock-in mice created by our group have strongly suggested that the farnesyl lipid modification may be critical for certain progeroid disorders, but less critical for others. We plan to explore the cell biology underlying this finding.

Public Health Relevance

The proteins of the nuclear lamina have generated enormous interest because of recent studies showing that genetic defects in nuclear lamins cause multiple human diseases. The main objective of this renewal application is to continue to explore the physiologic and medical relevance of the posttranslational processing of the nuclear lamins, particularly as it relates to devastating neuronal migration defects and the pathogenesis and treatment of progeroid syndromes (precocious aging syndromes). Our studies will be focused on the physiologic importance of the B-type lamins in the developing brain and other tissues and the physiologic importance of the posttranslational processing of prelamin A.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG035626-10
Application #
8284357
Study Section
Nuclear Dynamics and Transport (NDT)
Program Officer
Velazquez, Jose M
Project Start
2003-08-15
Project End
2014-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
10
Fiscal Year
2012
Total Cost
$400,013
Indirect Cost
$140,264
Name
University of California Los Angeles
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Lee, John M; Nobumori, Chika; Tu, Yiping et al. (2016) Modulation of LMNA splicing as a strategy to treat prelamin A diseases. J Clin Invest 126:1592-602
Yang, Shao H; Procaccia, Shiri; Jung, Hea-Jin et al. (2015) Mice that express farnesylated versions of prelamin A in neurons develop achalasia. Hum Mol Genet 24:2826-40
Tapia, Olga; Fong, Loren G; Huber, Michael D et al. (2015) Nuclear envelope protein Lem2 is required for mouse development and regulates MAP and AKT kinases. PLoS One 10:e0116196
Young, Stephen G; Jung, Hea-Jin; Lee, John M et al. (2014) Nuclear lamins and neurobiology. Mol Cell Biol 34:2776-85
Lee, John M; Tu, Yiping; Tatar, Angelica et al. (2014) Reciprocal knock-in mice to investigate the functional redundancy of lamin B1 and lamin B2. Mol Biol Cell 25:1666-75
Jung, Hea-Jin; Tu, Yiping; Yang, Shao H et al. (2014) New Lmna knock-in mice provide a molecular mechanism for the 'segmental aging' in Hutchinson-Gilford progeria syndrome. Hum Mol Genet 23:1506-15
Khan, Omar M; Akula, Murali K; Skålen, Kristina et al. (2013) Targeting GGTase-I activates RHOA, increases macrophage reverse cholesterol transport, and reduces atherosclerosis in mice. Circulation 127:782-90
Young, Stephen G; Yang, Shao H; Davies, Brandon S J et al. (2013) Targeting protein prenylation in progeria. Sci Transl Med 5:171ps3
Ibrahim, Mohamed X; Sayin, Volkan I; Akula, Murali K et al. (2013) Targeting isoprenylcysteine methylation ameliorates disease in a mouse model of progeria. Science 340:1330-3
Jung, Hea-Jin; Lee, John M; Yang, Shao H et al. (2013) Nuclear lamins in the brain - new insights into function and regulation. Mol Neurobiol 47:290-301

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