Abstract

Certain identifiable neurons and muscles of the moth Manduca sexta undergo a rapid, developmentally programmed cell death during metamorphosis to the adult. The commitment of these muscles to degenerate requires new RNA and protein synthesis. The present work seeks to identify genes which encode """"""""cell death"""""""" specific proteins. Poly A+ RNA will be used to construct stage-specific subtraction cDNA libraries in lambda GT10, and individual recombinants will be screened to confirm that they contain developmentally regulated cDNA sequences. These clones will then be sequenced to determine the primary structure of their presumptive protein products. Comparison of these sequences to those in protein databases may allow us to identify the nature of their function. Clones which appear to meet several defined criteria for cell death genes will be expressed in bacteria or antibody production. Both antibody and nucleic acid probes will be used to begin characterization of the mechanisms involved in regulating cell death during development in Manduca and a related moth species Antheraea polyphemus. The expression of homologous cell death genes will also be examined in frog and chicken using the antibodies and nucleic acid probes from Manduca.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM040458-03
Application #
3298008
Study Section
Molecular Cytology Study Section (CTY)
Project Start
1988-06-01
Project End
1991-05-31
Budget Start
1990-06-01
Budget End
1991-05-31
Support Year
3
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
University of Massachusetts Amherst
Department
Type
Schools of Arts and Sciences
DUNS #
153223151
City
Amherst
State
MA
Country
United States
Zip Code
01003

  Publications

Valavanis, Christos; Wang, Zhaohui; Sun, Danhui et al. (2007) Acheron, a novel member of the Lupus Antigen family, is induced during the programmed cell death of skeletal muscles in the moth Manduca sexta. Gene 393:101-9
Gao, Zheng-Liang; Deblis, Ryan; Glenn, Honor et al. (2007) Differential roles of HIC-5 isoforms in the regulation of cell death and myotube formation during myogenesis. Exp Cell Res 313:4000-14
Gao, Zhengliang; Schwartz, Lawrence M (2005) Identification and analysis of Hic-5/ARA55 isoforms: Implications for integrin signaling and steroid hormone action. FEBS Lett 579:5651-7
Wing, John P; Karres, Janina S; Ogdahl, Justyne L et al. (2002) Drosophila sickle is a novel grim-reaper cell death activator. Curr Biol 12:131-5
Schwartz, L M; Nambu, J R; Wang, Z (2002) Parkinsonism proteolysis and proteasomes. Cell Death Differ 9:479-82
Shumway, L; Schwartz, L M (2001) Generalized 96-well format for quantitative and qualitative monitoring of altered protein expression and posttranslational modification in cells. Biotechniques 31:996, 998, 1000
Kuelzer, F; Kuah, P; Bishoff, S T et al. (1999) Cloning and analysis of small cytoplasmic leucine-rich repeat protein (SCLP), a novel, phylogenetically-conserved protein that is dramatically up-regulated during the programmed death of moth skeletal muscle. J Neurobiol 41:482-94
Barnes, N Y; Li, L; Yoshikawa, K et al. (1998) Increased production of amyloid precursor protein provides a substrate for caspase-3 in dying motoneurons. J Neurosci 18:5869-80
Zhou, L; Schnitzler, A; Agapite, J et al. (1997) Cooperative functions of the reaper and head involution defective genes in the programmed cell death of Drosophila central nervous system midline cells. Proc Natl Acad Sci U S A 94:5131-6
Hashimoto, M K; Mykles, D L; Schwartz, L M et al. (1996) Imaginal cell-specific accumulation of the multicatalytic proteinase complex (proteasome) during post-embryonic development in the tobacco hornworm, Manduca sexta. J Comp Neurol 365:329-41

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