Download 슬라이드 1

Promoter activity of LTR element of the human FPRL2 gene
Hong-seok HA, Jae-Won HUH, Dae-Soo KIM1, Heui-Soo KIM *
Division of Biological Sciences, College of Natural Sciences, Pusan National University.
1PBBRC,
Interdisciplinary Research Program of Bioinformatics, College of Natural Sciences, Pusan National University.
HTTP://WWW.PRIMATE.OR.KR
ABSTRACT
INTRODUCTION
LTR: Long Terminal Repeat
The human genome is estimated to consist of approximately 8% human
endogenous retroviruses (HERVs) and related sequences. FPRL2 (fomyl peptide
receptor-like 2) gene has a solitary LTR (long terminal repeat). The LTR is
located between first exon and promoter region of the FPRL2 gene. The FPRL2
gene containing LTR element was expressed in various human tissues except
fetal brain and cerebellum. The LTR element was detected in hominoid, Old
World monkeys, and New World monkeys except for common marmoset,
whereas LINE (long interspersed repetitive element) and SINE (short interspersed
repetitive element) elements were detected in prosimian (ring-tailed lemur) and
common marmoset. We also examined promoter activity of the LTR element in
FPRL2 gene, and discussed its biological role. Taken together, the insertion of
retroelements into primate genome could have different biological roles during
primate evolution.
.
MATERIALS & METHODS
Bioinformatics
Luciferase Assay
Transfac 6.0
Real Time RT-PCR
Bisulfite Sequencing PCR
Genomic DNA PCR
& Gene cloning
LINE: Long Integrated nuclear element
SINE: Short Integrated nuclear element
SINE
13%
Other region
16%
Retroelement
HERV: Human Endogenous Retro-Virus
RNA intermediate
LINE
20%
- LTR element
+ LTR element
Retroposon
- env
Gene-related Sequence
36%
HERV element
8%
Retrotransposon
- RT
DNA element
3%
Pseudogene
Coding sequence
1%
+ RT
LTR
SINE
ORF1
ORF2
LTR
Yeast Ty1/copia/truncated HERVs
3%
LTR
P
LTR
Human THE1
Poly(A)
Human Alu
Retroelements have been subjected to
many amplification and transposition
events resulting in a widespread
distribution of complete or partial retroviral
sequences throughout the human genome.
The human genome comprises
approximately 8% of the human
endogenous retroviruses (HERVs) and
other long terminal repeat (LTR)–like
elements . Most HERVs seem to have
entered the genome between 10 and 50
million years ago, and they comprise over
200 distinct groups and subgroups .
Expression of retroelements can influence
the outcome of infections in different ways
that can be either beneficial or detrimental
to the host. A function of the multiple
copy families, scattered throughout the
genome, has been reported regulatory
functions on the gene expression of
nearby located genes . A small minority of
such sequences has acquired a role in
regulating gene expression, and some of
these may be related to differences
between individuals, and to expression of
disease.
.
+ env
Retrovirus
LINE
LTR
P
ORF1
ORF2
L1
Poly(A)
gag
pol
env
LTR
Full-length HERVs/exogenous retrovirus
REFERENCES
1. Maksakova IA, Mager DL. 2005. Transcriptional regulation of early transposon elements, an active family of mouse long terminal repeat
re t rot ra nsposons . J Vi ro .
.
2. Yang D, Chen Q, Gertz B, He R, Phulsuksombati M, Ye RD, Oppenheim JJ. 2002. Human dendritic cells express functional formyl peptide
receptor-like-2 (FPRL2) throughout maturation. J Leukoc Biol.
.
RESULTS & DISCUSSION
Fig. 1. The genomic structure of FPRL2 gene including LTR54 element. Exons were represented by solid box
with the exon numbers. Arrows indicate the primer location. The LTR54 element was integrated into the
FPRL2 gene with the antisense orientation on human chromosome 19q13.41
Fig. 4. PCR analysis for the presence of LTR54-derived promoter region of FPRL2 gene using the various primate
genomic DNAs (A). Primates showed PCR products that were cloned and sequenced (B,C).
Fig. 2. Real Time RT-PCR analysis of LTR54 derived transcript from different human tissues.
Fig. 3. Luciferase reporter gene assay for LTR54-derived promoter of FPRL2 gene in transient transfected
HCT116. Relative activity of luciferase assay for pGL2-hFPRL2-LTR54 in forward and reverse orientation or the
pGL2 basic vector was indicated as schematic diagram. Results are expressed as ratios of the luciferase activity to
that of the promoterless pGL2 reporter plasmid.
Fig 5. Comparison of FPRL2 gene family in silico analysis; gene location (A), comparison of mRNA
structure using PIP maker program(B) , alignment of amino acid (C)