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Conserved overlapping gene arrangement, restricted expression, and biochemical activities of DNA polymerase ν (POLN).

Takata K, Tomida J, Reh S, Swanhart LM, Takata M, Hukriede NA, Wood RD - J. Biol. Chem. (2015)

Bottom Line: Consistent with this, injection of POLN-specific morpholino antisense oligonucleotides did not interfere with zebrafish embryonic development.These properties are conserved with the human enzyme.Although the physiological function of pol ν remains to be clarified, this study uncovers distinctive aspects of its expression control and evolutionarily conserved properties of this DNA polymerase.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Epigenetics and Molecular Carcinogenesis, University of Texas M.D. Anderson Cancer Center, Smithville, Texas 78957, the University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas 77030, ktakata@mdanderson.org.

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A, sequence alignment of 5′-RACE products for the human POLN cDNA. The newly identified 5′-untranslated region is shown within the box. A well matched Kozak consensus sequence for translation initiation surrounds the ATG start codon. B, sequence alignment of the first exon of human POLN (sequence of RACE-PCR clone B7 shown in A) and HAUS3 mRNA (NCBI reference sequence, NM_024511.5), and mouse Poln (AY135562) and Haus3 (NM_146159). Perfectly aligned sequences are boxed. The precise transcription initiation site is not known for POLN or HAUS3 and could extend slightly 5′ of that shown for both mouse and human mRNAs.
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Figure 3: A, sequence alignment of 5′-RACE products for the human POLN cDNA. The newly identified 5′-untranslated region is shown within the box. A well matched Kozak consensus sequence for translation initiation surrounds the ATG start codon. B, sequence alignment of the first exon of human POLN (sequence of RACE-PCR clone B7 shown in A) and HAUS3 mRNA (NCBI reference sequence, NM_024511.5), and mouse Poln (AY135562) and Haus3 (NM_146159). Perfectly aligned sequences are boxed. The precise transcription initiation site is not known for POLN or HAUS3 and could extend slightly 5′ of that shown for both mouse and human mRNAs.

Mentions: Exons encoding the HAUS3 protein are located just upstream of and closely adjacent to POLN (Fig. 2). HAUS3 encodes subunit 3 of the multisubunit augmin protein complex, critical for regulation of centrosome and spindle integrity. This syntenic relationship appears to be conserved in vertebrates. Surprisingly, our searches of transcript databases revealed that the zebrafish POLN transcript overlaps with the 5′UTR of HAUS3, indicating that both share the first exon (Fig. 2A). To investigate this further, we used RACE-PCR to determine the 5′UTR sequence of human POLN. All of the 5′UTR sequence clones for HsPOLN identified in this analysis overlapped with the 5′UTR of HsHAUS3 (Fig. 3A). Similarly, the 5′UTR of the Poln transcript isolated from mouse testis RNA (2) overlaps with the 5′UTR sequence of MmHaus3 (Fig. 3B). Thus for zebrafish, human, and mouse POLN, experimental data show that the coding exons for another gene (HAUS3) reside within the first intron (Fig. 2B), a unique arrangement among DNA polymerases.


Conserved overlapping gene arrangement, restricted expression, and biochemical activities of DNA polymerase ν (POLN).

Takata K, Tomida J, Reh S, Swanhart LM, Takata M, Hukriede NA, Wood RD - J. Biol. Chem. (2015)

A, sequence alignment of 5′-RACE products for the human POLN cDNA. The newly identified 5′-untranslated region is shown within the box. A well matched Kozak consensus sequence for translation initiation surrounds the ATG start codon. B, sequence alignment of the first exon of human POLN (sequence of RACE-PCR clone B7 shown in A) and HAUS3 mRNA (NCBI reference sequence, NM_024511.5), and mouse Poln (AY135562) and Haus3 (NM_146159). Perfectly aligned sequences are boxed. The precise transcription initiation site is not known for POLN or HAUS3 and could extend slightly 5′ of that shown for both mouse and human mRNAs.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591814&req=5

Figure 3: A, sequence alignment of 5′-RACE products for the human POLN cDNA. The newly identified 5′-untranslated region is shown within the box. A well matched Kozak consensus sequence for translation initiation surrounds the ATG start codon. B, sequence alignment of the first exon of human POLN (sequence of RACE-PCR clone B7 shown in A) and HAUS3 mRNA (NCBI reference sequence, NM_024511.5), and mouse Poln (AY135562) and Haus3 (NM_146159). Perfectly aligned sequences are boxed. The precise transcription initiation site is not known for POLN or HAUS3 and could extend slightly 5′ of that shown for both mouse and human mRNAs.
Mentions: Exons encoding the HAUS3 protein are located just upstream of and closely adjacent to POLN (Fig. 2). HAUS3 encodes subunit 3 of the multisubunit augmin protein complex, critical for regulation of centrosome and spindle integrity. This syntenic relationship appears to be conserved in vertebrates. Surprisingly, our searches of transcript databases revealed that the zebrafish POLN transcript overlaps with the 5′UTR of HAUS3, indicating that both share the first exon (Fig. 2A). To investigate this further, we used RACE-PCR to determine the 5′UTR sequence of human POLN. All of the 5′UTR sequence clones for HsPOLN identified in this analysis overlapped with the 5′UTR of HsHAUS3 (Fig. 3A). Similarly, the 5′UTR of the Poln transcript isolated from mouse testis RNA (2) overlaps with the 5′UTR sequence of MmHaus3 (Fig. 3B). Thus for zebrafish, human, and mouse POLN, experimental data show that the coding exons for another gene (HAUS3) reside within the first intron (Fig. 2B), a unique arrangement among DNA polymerases.

Bottom Line: Consistent with this, injection of POLN-specific morpholino antisense oligonucleotides did not interfere with zebrafish embryonic development.These properties are conserved with the human enzyme.Although the physiological function of pol ν remains to be clarified, this study uncovers distinctive aspects of its expression control and evolutionarily conserved properties of this DNA polymerase.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Epigenetics and Molecular Carcinogenesis, University of Texas M.D. Anderson Cancer Center, Smithville, Texas 78957, the University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas 77030, ktakata@mdanderson.org.

Show MeSH
Related in: MedlinePlus