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The nucleosome as a potential barrie...

Golding, Amit.

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The nucleosome as a potential barrier to the V(D)J recombinase.

紀錄類型:	書目-語言資料,印刷品 : Monograph/item
正題名/作者:	The nucleosome as a potential barrier to the V(D)J recombinase./
作者:	Golding, Amit.
面頁冊數:	135 p.
附註:	Advisers: Mark Schlissel; Stephen Desiderio.
Contained By:	Dissertation Abstracts International63-03B.
標題:	Biology, Genetics. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3046459
ISBN:	0493606491

The nucleosome as a potential barrier to the V(D)J recombinase.
Golding, Amit.

The nucleosome as a potential barrier to the V(D)J recombinase. - 135 p.

Advisers: Mark Schlissel; Stephen Desiderio.

Thesis (Ph.D.)--The Johns Hopkins University, 2002.

Lineage specificity and temporal ordering of immunoglobulin (Ig) and T cell receptor (TCR) gene rearrangement are reflected in the accessibility of recombination signal sequences (RSSs) within chromatin to in vitro cleavage by the V(D)J recombinase. In the present work, I investigated the basis of this regulation by testing the ability of purified RAG1 and RAG2 proteins to initiate cleavage on positioned nucleosomes containing RSS substrates. I found that nicking and double-strand DNA cleavage of RSSs positioned on the face of an unmodified nucleosome are entirely inhibited. This inhibition was independent of translational position or rotational phase and could not be overcome either by addition of the DNA-bending protein HMG-1 or by the use of hyperacetylated histones. I suggest that the nucleosome could act as the stable unit of chromatin that limits recombinase accessibility to potential RSS targets, and that actively rearranging gene segments might be packaged in a modified or disrupted nucleosome structure. Using techniques based on MNase and restriction enzyme digestion of intact nuclei, I examined the in vivo chromatin structure of the immunoglobulin kappa locus J genes in non-lymphoid cells as compared to lymphoid cells poised at different stages of development. I present preliminary data from general MNase sensitivity, indirect end-labeling, high resolution mapping of MNase breaks by LMPCR, and restriction enzyme accessibility.

ISBN: 0493606491Subjects--Topical Terms:

1017730
Biology, Genetics.

The nucleosome as a potential barrier to the V(D)J recombinase.
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502 $a Thesis (Ph.D.)--The Johns Hopkins University, 2002.
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