PMID- 8593612
OWN - NLM
STAT- MEDLINE
DCOM- 19960409
LR  - 20191210
IS  - 1088-9051 (Print)
IS  - 1088-9051 (Linking)
VI  - 5
IP  - 3
DP  - 1995 Oct
TI  - Functional expression of yeast artificial chromosome-human multidrug resistance 
      genes in mouse cells.
PG  - 245-58
AB  - Multidrug resistance (MDR) genes, which are ATP-binding cassette family genes, 
      encode the cell surface glycoprotein, P-glycoprotein, which functions as an 
      energy-dependent drug efflux pump. Two relevant human genes, PGY1 and PGY3, are 
      located on human chromosome 7, and three relevant mouse genes, mdr1a, mdr1b, and 
      mdr2, are located on mouse chromosome 5. An LMD1 cell line was established after 
      the transfer of a 580-kb yeast artificial chromosome (YAC) clone carrying the 
      human MDR locus into mouse L cells; the cell line was shown to have stably 
      integrated YAC DNA in an apparent intact form. Using LMD1 cells as the parental 
      cell line, five vincristine-resistant sublines, designated LMD1-V50, LMD1-V100, 
      LMD1-V200, LMD1-V500, and LMD1-V1000, were isolated by exposure to increasing 
      concentrations of the drug. LMD1-V50, LMD1-V100, LMD1-V200, LMD1-V500, and 
      LMD1-V1000 showed 3-, 7-, 13-, 45-, and 110-fold higher resistance to the 
      cytotoxic effects of vincristine, respectively, than their parental counterpart, 
      LMD1. Immunofluorescence, Western blot, and Northern blot analyses revealed that 
      the human PGY1 gene or its product was overexpressed, accompanied by gene 
      amplification. The human PGY3 gene was also overexpressed in the LMD1-V20, 
      LMD1-V100, and LMD1-V1000 cell lines. Southern blot and fluorescence in situ 
      hybridization (FISH) analyses demonstrated that although essentially the entire 
      YAC DNA was integrated in mouse genome and amplified, the endogenous mouse mdr 
      genes were not amplified in these drug-resistant cell lines. Similar results were 
      obtained by the analyses of vincristine-resistant cell lines isolated from four 
      independent subclones of LMD1 cells. Thus, in contrast to their mouse 
      counterparts, the integrated human MDR genes retained susceptibility to both gene 
      activation and amplification, during the selection of drug-resistant mouse cell 
      lines. The possibility that transferred YACs may retain regulatory properties 
      observed in the cells of origin, and may have a chromatin structure that favors 
      augmented expression, is discussed.
FAU - Kusaba, H
AU  - Kusaba H
AD  - Department of Biochemistry, Kyushu University School of Medicine, Fukuoka, Japan.
FAU - Kohno, K
AU  - Kohno K
FAU - Asakuno, K
AU  - Asakuno K
FAU - Kuwano, M
AU  - Kuwano M
FAU - Okumura, K
AU  - Okumura K
FAU - Green, E D
AU  - Green ED
FAU - Schlessinger, D
AU  - Schlessinger D
FAU - Wada, M
AU  - Wada M
LA  - eng
GR  - P50-HG00201/HG/NHGRI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Genome Res
JT  - Genome research
JID - 9518021
RN  - 0 (ATP Binding Cassette Transporter, Subfamily B)
RN  - 0 (ATP Binding Cassette Transporter, Subfamily B, Member 1)
RN  - 0 (ATP-Binding Cassette Transporters)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 5J49Q6B70F (Vincristine)
RN  - 9EI49ZU76O (multidrug resistance protein 3)
SB  - IM
MH  - *ATP Binding Cassette Transporter, Subfamily B
MH  - ATP Binding Cassette Transporter, Subfamily B, Member 1/biosynthesis/*genetics
MH  - ATP-Binding Cassette Transporters/biosynthesis/*genetics
MH  - Animals
MH  - Base Sequence
MH  - Blotting, Western
MH  - Chromosomes, Artificial, Yeast
MH  - Chromosomes, Human, Pair 7
MH  - Drug Resistance, Multiple/*genetics
MH  - Drug Resistance, Neoplasm/genetics
MH  - Gene Expression
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - KB Cells/drug effects/metabolism
MH  - L Cells/drug effects/metabolism
MH  - Mice
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction
MH  - Recombinant Fusion Proteins/biosynthesis/genetics
MH  - Vincristine/pharmacology
EDAT- 1995/10/01 00:00
MHDA- 1995/10/01 00:01
CRDT- 1995/10/01 00:00
PHST- 1995/10/01 00:00 [pubmed]
PHST- 1995/10/01 00:01 [medline]
PHST- 1995/10/01 00:00 [entrez]
AID - 10.1101/gr.5.3.245 [doi]
PST - ppublish
SO  - Genome Res. 1995 Oct;5(3):245-58. doi: 10.1101/gr.5.3.245.