Bayir, Erkan
(2016)
ROLE OF HNRNP H IN A REVERSIBLE FINE-TUNING MECHANISM OF
SPLICING RESPONSE UPON TRANSCRIPTIONAL PAUSING.
Master's Thesis, University of Pittsburgh.
(Unpublished)
This is the latest version of this item.
Abstract
Alternative splicing is a common way of diversifying the proteome and regulating protein functions without increasing genome size. Chemotherapeutic agents, such as cisplatin, cause changes in the splicing of proliferation-associated exons, which contribute to the overall proliferative phenotype of cancer cells. Transcriptional pausing, which is seen in embryonic stem cells as a timing regulator of gene expression, also impacts splicing. The mechanisms by which transcriptional pausing and pause-release alter the functions of splicing machinery are largely unknown. Thus, I tested the effects of the transcription elongation inhibitor DRB (5,6-dichloro-1-β-D-ribofuranosylbenzimidazole) on the splicing of a set of inducible exons with a common UAGG-GGGG splicing code. These exons were previously identified through a genomewide search for the transcripts sensitive to hnRNP A1 and hnRNP H. I found that the endogenous human HNRNPH1 Exon 4 and its paralog HNRNPH3 Exon 3 had increased skipping upon DRB treatment, as did the pA+ reporter-cloned GRIN1 CI, a brain-region-specific rat exon with this splicing code, previously studied in our lab. I used single and combined mutations to monitor the effect of DRB-induced transcriptional pausing and pause-release on splicing, since the shared motifs include two exonic UAGG motifs and a GGGG close to 5´ splice site. GGGG mutants had increased skipping, compared to wild type, upon DRB treatment. UAGG-GGGG code is necessary for the response, as the triple mutant lost its ability to respond to DRB. These data point to a role of hnRNP H in response to transcription stress.
Share
| Citation/Export: |
|
| Social Networking: |
|
Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
|
| ETD Committee: |
|
| Date: |
20 September 2016 |
| Date Type: |
Publication |
| Defense Date: |
7 July 2016 |
| Approval Date: |
20 September 2016 |
| Submission Date: |
1 August 2016 |
| Access Restriction: |
No restriction; Release the ETD for access worldwide immediately. |
| Number of Pages: |
119 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
Dietrich School of Arts and Sciences > Biological Sciences |
| Degree: |
MS - Master of Science |
| Thesis Type: |
Master's Thesis |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
splicing, transcription, stress, pause-release, DRB |
| Date Deposited: |
20 Sep 2016 15:37 |
| Last Modified: |
15 Nov 2016 14:35 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/29176 |
Available Versions of this Item
-
ROLE OF HNRNP H IN A REVERSIBLE FINE-TUNING MECHANISM OF
SPLICING RESPONSE UPON TRANSCRIPTIONAL PAUSING. (deposited 20 Sep 2016 15:37)
[Currently Displayed]
Metrics
Monthly Views for the past 3 years
Plum Analytics
Actions (login required)
 |
View Item |
![[feed]](/images/feed-icon-32x32.png){kind=icon}