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Study of the molecular mechanism of the Neurospora circadian clock.

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Call No.	QH470.N4 L86 1998
Author	Luo, Chenghua
Title	Study of the molecular mechanism of the Neurospora circadian clock.
Date	1998
Abstract	The frequency gene and the two proteins it encodes are central components of the Neurospora circadian clock. The frq message and proteins are involved in a negative feedback loop in which FRO represses its own transcript level. Based on this feedback loop, a genetic screen was designed and performed to search for other components involved in this repression. A total of 1.6 x 10 5 independent mutants were screened, 17 putative clock mutants were isolated. All 17 mutants were analyzed by crosses with strain frq7 un-10 (732-31). A minimum of 33 progeny were tested from each cross. Correct segregation of the different loci suggested that the crosses were normal but none of the mutations was associated with the clock. Positive results have been obtained from each step of this mutant screen; only the final result is negative in that no new clock mutants have been identified. This result suggests that the feedback loop may be assembled differently from what has been expected. In order to study the molecular mechanism of the FRQ protein during the circadian cycle, a biochemical approach was used. The subcellular localization of FRQ was determined by separating the nuclear fractions on SDS-PAGE followed by western blot. The results demonstrate that FRQ is a nuclear protein, that it cycles inside the nucleus, and that its nuclear cycle which is similar to that of the frq mRNA cycle is phase advanced with respect to that of the total FRQ cycle. This correlates well with data showing that FRQ executes its negative effect on its transcript''s level within three to six hours of synthesis, suggesting a possible mechanism for how FRQ may achieve its negative effect on its message level. Results showed that FRQ's nuclear localization is essential for its function in the clock. Further experiments attempting to address FRQ's ability to bind DNA are also presented.
Description	1 online resource (xii, 176 leaves) : illustrations
Format	application/pdf
Advisor	Taylor, Ron K.
Subject	Neurospora -- Genetics. ; Neurospora -- Effect of light on. ; Circadian rhythms. ; Proteins -- Synthesis -- Effect of light on. ; Genetic regulation.
College	Dartmouth Medical School -- Department of Biochemistry.
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.2807

Title	luo1998 1
Full Text	Study of the Molecular Mechanism of the Neurospora Circadian Clock A Thesis Submitted to the Faculty in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biochemistry by Chenghua Luo Dartmouth College Hanover, NH 03755 October, 1998 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.

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