Unit:
Τομέας Βιοχημείας Μοριακής ΒιολογίαςLibrary of the School of Science
Author:
Καρούσης Ευάγγελος
Dissertation committee:
Αναπλ. Καθηγητής Διαμάντης Σίδερης (επιβλέπων), Καθηγητής Εμμανουήλ Φραγκούλης, Ερευνητής Α' Γεώργιος Παναγιώτου
Original Title:
Μελέτη της έκφρασης της ανθρώπινης ριβονουκλεάσης κ και πιθανών ισομορφών της σε διάφορους ιστούς
Translated title:
Expression analysis of human RNase κ in several tissues
Summary:
Subtle alternative splicing leads to the formation of RNA variants lacking or
including a small number of nucleotides. To date, the impact of subtle
alternative splicing phenomena on protein biosynthesis has been studied in
frame-preserving incidents. On the contrary, mRNA isoforms derived from
frame-shifting events were poorly studied and generally characterized as
non-coding. This work provides evidence for a frame-shifting subtle alternative
splicing event which results in the production of a novel protein isoform. We
applied a combined molecular approach for the cloning and expression analysis
of a human RNase κ transcript (RNase κ-02) which lacks four consecutive bases
compared to the previously isolated RNase κ isoform. RNase κ-02 mRNA is
expressed in all human cell lines tested end encodes the synthesis of a
134-amino-acid protein by utilizing an alternative initiation codon. The
expression of RNase κ-02 in the cytoplasm of human cells was verified by
Western blot and immunofluorescence analysis using a specific polyclonal
antibody developed on the basis of the amino-acid sequence difference between
the two protein isoforms. The results presented here show that subtle changes
during mRNA splicing can lead to the expression of significantly altered
protein isoforms.
Keywords:
Ribonuclease, Alternative splicing, Subtle alternative splicing, Gene expression, Biochemical characterization
Number of references:
185
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