Find results with:

Add another field

Search by date

to

Searching collections:

Dartmouth Dissertations

Add or remove collections

Home Dartmouth Dissertations krieser1999 1

Reference URL

To link to the entire object, paste this link in email, IM or document

To embed the entire object, paste this HTML in website

To link to this page, paste this link in email, IM or document

To embed this page, paste this HTML in website

The study of an endonuclease and its potential role in apoptosis.

you wish to report:

Your comment:

Your Name:

Submit

Cancel

...

Back to top

Powered by CONTENTdm®

Call No.	QP609.D4 K74 1999
Author	Krieser, Ronald
Title	The study of an endonuclease and its potential role in apoptosis.
Date	1999
Abstract	Previous work in this laboratory has implicated deoxyribonuclease II (DNase II) as an endonuclease responsible for DNA digestion during apoptosis. The goal of this thesis was to isolate the cDNA encoding DNase II and study its potential role in apoptosis. The cDNA contains an open reading frame of 1078 bases encoding a 40 kDa protein, although the mature protein is 30 kDa. The gene was localized to chromosome 19p13.2, and has no significant homology to other human genes or proteins. Genbank database searches identified three C. elegans genes encoding "putative proteins" with 31-36% identity to the human endonuclease. Transfection of the human DNase II cDNA into Chinese hamster ovary cells induced chromatin condensation, a hallmark of apoptosis. This chromatin condensation was not prevented by inhibitors of apoptosis demonstrating the effect was directly due to DNase II rather than through activation of alternate apoptosis effector molecules. Our attempts to raise an anti-DNase II antibody failed, which may be explained by the high homology of DNase II observed between species and the normal presence of DNase II in plasma which may lead to immunotolerance of the antigen. However, one antisera recognized a protein that was specifically cleaved from a 26 kDa form to a 22 kDa form. This protein was purified and sequenced and identified as D4-GDI. Additional studies showed that the 26 kDa D4-GDI resided in the cytoplasm of undamaged cells, whereas following cleavage, the 22 kDa form translocated to the nucleus. Mutations created in D4-GDI confirmed cleavage at a caspase 3 site, yet cleavage-resistant mutants did not suppress apoptosis demonstrating that cleavage may not be required for apoptosis. This is the first report describing translocation of a caspase substrate from the cytosol to the nucleus after proteolysis. In summary, the results with DNase II continue to implicate it as an endonuclease potentially involved in apoptosis.
Description	1 online resource (xii, 128 leaves) : illustrations
Format	application/pdf
Advisor	Eastman, Alan
Subject	Deoxyribonucleases. ; Endonucleases. ; Apoptosis.
College	Dartmouth Medical School -- Department of Pharmacology and Toxicology.
Issuing Body	Dartmouth Dissertations are made available through the Dartmouth Digital Library.
Degree	Thesis (Ph.D)--Dartmouth College.
Copyright	In Copyright. This material is protected by copyright and/or related rights. You are free to use this material in any way that is permitted by copyright law that applies to your use. For other uses, you need to obtain permission from the rights-holder(s). http://rightsstatements.org/page/InC/1.0/
DOI	https://doi.org/10.1349/ddlp.3199

Title	krieser1999 1
Full Text	The Study of an Endonuclease and its Potential Role in Apoptosis A Thesis Submitted to the Faculty in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Pharmacology and Toxicology by Ronald Krieser DARTMOUTH COLLEGE Hanover, New Hampshire January 20, 1999 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.

Find results with:

Searching collections:

Home Dartmouth Dissertations krieser1999 1

The study of an endonuclease and its potential role in apoptosis.

Object Description

Description

The study of an endonuclease and its potential role in...

The study of an endonuclease and its potential role in...

	All
	Pages Enter page numbers or page ranges separated by commas. For example: 1,3,5-12