. Embedment-free electron microscopy shows the nuclear matrix comprised of a network of 11 nm filaments with numerous embedded dense organelles. They will use two new nuclear matrix preparations, high salt and the stabilized pre- fixed matrix. They will determine the true nature of the interchromatin granules using reinless section EM of the nuclear matrix preparations immunostained with antibodies to splicing components (in collaboration with Dr. Phil Sharp laboratory). They will combine these results with in situ hybridization with intron specific probes (in collaboration with Dr.Jeanne Lawrence) to locate which nuclear matrix organelle(s) are the location of splicing. In situ hybridization of hnRNA tracks will image the RNA transport machinery. Hybridization with an intermediate repetitive sequence will identify the location of hnRNA that does not serve as mRNA precursor. They will seek monoclonal antibodies to NM filament proteins to determine their number, molecular sequences and fate at mitosis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA067628-34
Application #
6172768
Study Section
Special Emphasis Panel (ZRG1-SSS-I (02))
Program Officer
Mohla, Suresh
Project Start
1995-06-15
Project End
2002-03-31
Budget Start
2000-04-01
Budget End
2001-03-31
Support Year
34
Fiscal Year
2000
Total Cost
$366,675
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139
Wan, K M; Nickerson, J A; Krockmalnic, G et al. (1999) The nuclear matrix prepared by amine modification. Proc Natl Acad Sci U S A 96:933-8
Penman, J; Penman, S (1997) Resinless section electron microscopy reveals the yeast cytoskeleton. Proc Natl Acad Sci U S A 94:3732-5
Nickerson, J A; Krockmalnic, G; Wan, K M et al. (1997) The nuclear matrix revealed by eluting chromatin from a cross-linked nucleus. Proc Natl Acad Sci U S A 94:4446-50
Penman, S (1995) Rethinking cell structure. Proc Natl Acad Sci U S A 92:5251-7