In the first part of the proposed studies we will examine further the mechanisms through which nutritional state regulates apo Beta synthesis and VLDL assembly. We will examine the mechanism through which sucrose feeding increases apo Beta secretion without affecting the hepatic content of apo Beta mRNA. We will test our hypothesis that sucrose feeding increases VLDL assembly by increasing the portion of apo Beta which enters the VLDL assembly pathway. Secondly, we will examine the mechanism through which fasting selectively decreases apo Beta synthesis by decreasing apo Beta mRNA concentrations. We will determine absolute rates of apo Beta mRNA transcription and degradation in both control and fasted rats. In the second part of the proposed studies we will further examine our hypothesis that integration into the endoplasmic reticular and binding to actin affords apo Beta a mechanism through which triglyceride can be concentrated in the endoplasmic reticulum and combined into a VLDL particle which buds off into the lumen in a manner similar to viruses. To examine this hypothesis further, specific sequences of human apo Beta100 cDNA shown by computer analysis to contain a putative actin binding domain, putative hydrophobic stop-transfer signals and the putative apo Beta48 junction will be inserted in-frame with a non-processed reporter protein (alpha-globin). The expression of the apo Beta-globin fusion proteins in different cell types having species, tissue and genetic specific traits will enable us to examine if the apo Beta sequences can make globin behave in a manner similar to apo Beta100 in regard to secretory processes expressed by these cells. The combined physiologic, cell and molecular biologic approaches should provide new insights into the mechanisms through which VLDL assembly is regulated.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL041624-03
Application #
3359446
Study Section
Nutrition Study Section (NTN)
Project Start
1988-12-01
Project End
1993-11-30
Budget Start
1990-12-15
Budget End
1991-11-30
Support Year
3
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Colorado Denver
Department
Type
Schools of Medicine
DUNS #
065391526
City
Aurora
State
CO
Country
United States
Zip Code
80045