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Determining the effects of ribonucleoprotein hnRNP D40 on the alternative splicing of HPV-16 E6/E7 and E1/E2 and identifying hnRNP D40’s nuclear export pathway

Ortís Sunyer, Júlia (2020) MOBN03 20192
Degree Projects in Molecular Biology
Popular Abstract
Papillomavirus, the leading cause of cervical cancer

Papillomaviruses are family of viruses that infect mammals and their uncured infection can lead to cancer. In humans, they can cause as much as 5% of all the cancers worldwide. Even though some papillomavirus types can also cause warts on hands and feet that will clear on its own, some types, regarded as high-risk by specialists, can infect the anogenital area and lead to cancer if not cleared. Even though the transmission has been reduced by both extensive campaigns of vaccination and by the use of sexual protective measures, there are so many things yet to discover about the virus’s regulation in the human cell.

Papillomaviruses use the infected cell’s machinery to obtain more... (More)
Papillomavirus, the leading cause of cervical cancer

Papillomaviruses are family of viruses that infect mammals and their uncured infection can lead to cancer. In humans, they can cause as much as 5% of all the cancers worldwide. Even though some papillomavirus types can also cause warts on hands and feet that will clear on its own, some types, regarded as high-risk by specialists, can infect the anogenital area and lead to cancer if not cleared. Even though the transmission has been reduced by both extensive campaigns of vaccination and by the use of sexual protective measures, there are so many things yet to discover about the virus’s regulation in the human cell.

Papillomaviruses use the infected cell’s machinery to obtain more copies of themselves. This brutal take of the cell by the virus is what can lead to a missregulation of the cell division and cause an uncontrolled growth. One of the things that the virus particles use for their benefit are heterogeneous ribonucleoproteins (hnRNP). HNRNPs are a type of proteins that bind to RNA and can influence its maturation and, therefore, influence the protein expression in the cells. One of their members is hnRNP D, which can bind to the papillomavirus genome and modify its protein expression. hnRNP D, which can be found in both nucleus and cytoplasm, has four different possible forms, each of which distinguished by its weight: 37, 40, 42 and 45 kDa.

In my study, various molecular biology techniques are used to elucidate how hnRNP D40 influences human papillomavirus type 16 (HPV-16) gene expression. The results show that hnRNP D40 inhibits the alternative splicing of the genes E6/E7 and E1/E2 leading to the production of the full-length transcripts. When hnRNP D40 is absent, certain shorter forms are produced instead. In addition, we also tried to show where hnRNP D40 was binding on HPV-16. At the moment, we have one possible binding site. Nevertheless, more experiments are required in order to properly elucidate them.

In this project, we also wanted to unveil which hnRNP D40’s regions are crucial to perform its function. It seems that when the start of the protein is missing, called N-terminal in scientific slang, hnRNP D40 can still maintain most of its functions. This leads us to believe that the end of the protein is of upmost importance to correctly develop its functions. Another study carried out in the laboratory was to test if some mutations on exon 2, found in this N-terminal domain, caused a change of hnRNP D40’s activity. Nevertheless, the results could not be taken into account due to bigger mutation problems in the cloned hnRNP D40’s DNA.

In addition to all this, we were set to know how hnRNP D40 was able to go from the nucleus to the cytoplasm of the cell. For this reason, the same molecular biology methods were used to determine if hnRNP D40 bound to TAP, a nuclear export protein, to go to the cytoplasm. The results, however, show that hnRNP D40 might have another way of shuttling between the two major cell compartments.

Master’s Degree Project in Molecular Biology 60 credits 2020
Department of Biology, Lund University

Advisor: Stefan Schwartz
HPV group, Laboratory medicine, BMC B13 (Less)
Please use this url to cite or link to this publication:
author
Ortís Sunyer, Júlia
supervisor
organization
course
MOBN03 20192
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
9028681
date added to LUP
2020-09-09 11:02:22
date last changed
2020-09-09 11:02:22
@misc{9028681,
  author       = {{Ortís Sunyer, Júlia}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Determining the effects of ribonucleoprotein hnRNP D40 on the alternative splicing of HPV-16 E6/E7 and E1/E2 and identifying hnRNP D40’s nuclear export pathway}},
  year         = {{2020}},
}