Cloning of a rat cDNA encoding a novel LIM domain protein with high homology to rat RIL

Haichao Wang; D. Corinne Harrison-Shostak; John J. Lemasters; Brian Herman

doi:10.1016/0378-1119(95)00542-E

Cloning of a rat cDNA encoding a novel LIM domain protein with high homology to rat RIL

Haichao Wang, D. Corinne Harrison-Shostak, John J. Lemasters, Brian Herman

Biomedical Engineering

Research output: Contribution to journal › Article › peer-review

46 Scopus citations

Abstract

A complementary DNA (1392 bp) encoding a protein with high homology to rat reversion-induced LIM (RIL) protein was cloned from rat hepatocytes by differential screening of a subtractive (normoxic minus hypoxic) λGEM-2 cDNA library. This cDNA clone, denoted CLP-36, encodes a 327-amino-acid (aa) protein that contained a restrictively conserved LIM (a Cys-rich domain with consensus aa sequence C-X₂-C-X_17-19-H-X₂-C-X₂-C-X₂-C-X_16-20-C-X₂-C/ H/D that was initially identified in homeodomain proteins, Lin-11 [Freyd et al., Nature 344 (1990) 876-879], Isl-1 [Karlsson et al., Nature 344 (1990) 879-882] and Mec-3 [Way et al., Cell 54 (1988) 5-16] in the C-terminal portion (aa 192-327). It shared an overall 45.1% identity to a rat LIM domain RIL protein [Kiess et al., Oncogene 10 (1995) 61-68], with 62.0% identity in the N terminus (aa 1-89) and 50.0% identity in the C terminus (aa 188-327). The rat CLP-36 mRNA is expressed most abundantly in heart, lung and liver, moderately in spleen and skeletal muscle, and at extremely low levels (if at all) in testis and brain tissues. Northern blotting analysis of total RNA extracted from normoxic or hypoxic rat hepatocytes, with a fragment of clone CLP-36 as probe, demonstrated a single band with a mobility corresponding to a size of 1.4 kb, whose level was significantly decreased during chemical hypoxia.

Original language	English (US)
Pages (from-to)	267-271
Number of pages	5
Journal	Gene
Volume	165
Issue number	2
DOIs	https://doi.org/10.1016/0378-1119(95)00542-E
State	Published - 1995

Bibliographical note

Funding Information:
We thank Dr. W. Litaker of the Biotechnology Center of University of North Carolina for his valuable discussions, and Drs. A. Periasamy, G. Gordon and P. Wodnicki for their expert assistance in using computer for DNA database analysis. This research was supported by grant AG07218 from NIH to B.H.

Keywords

Hepatocyte
PCR
RACE
amino-acid sequence
chemical hypoxia
nucleotide sequence
phage library
recombinant DNA

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10.1016/0378-1119(95)00542-E

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title = "Cloning of a rat cDNA encoding a novel LIM domain protein with high homology to rat RIL",

abstract = "A complementary DNA (1392 bp) encoding a protein with high homology to rat reversion-induced LIM (RIL) protein was cloned from rat hepatocytes by differential screening of a subtractive (normoxic minus hypoxic) λGEM-2 cDNA library. This cDNA clone, denoted CLP-36, encodes a 327-amino-acid (aa) protein that contained a restrictively conserved LIM (a Cys-rich domain with consensus aa sequence C-X2-C-X17-19-H-X2-C-X2-C-X2-C-X16-20-C-X2-C/ H/D that was initially identified in homeodomain proteins, Lin-11 [Freyd et al., Nature 344 (1990) 876-879], Isl-1 [Karlsson et al., Nature 344 (1990) 879-882] and Mec-3 [Way et al., Cell 54 (1988) 5-16] in the C-terminal portion (aa 192-327). It shared an overall 45.1% identity to a rat LIM domain RIL protein [Kiess et al., Oncogene 10 (1995) 61-68], with 62.0% identity in the N terminus (aa 1-89) and 50.0% identity in the C terminus (aa 188-327). The rat CLP-36 mRNA is expressed most abundantly in heart, lung and liver, moderately in spleen and skeletal muscle, and at extremely low levels (if at all) in testis and brain tissues. Northern blotting analysis of total RNA extracted from normoxic or hypoxic rat hepatocytes, with a fragment of clone CLP-36 as probe, demonstrated a single band with a mobility corresponding to a size of 1.4 kb, whose level was significantly decreased during chemical hypoxia.",

keywords = "Hepatocyte, PCR, RACE, amino-acid sequence, chemical hypoxia, nucleotide sequence, phage library, recombinant DNA",

author = "Haichao Wang and Harrison-Shostak, {D. Corinne} and Lemasters, {John J.} and Brian Herman",

note = "Funding Information: We thank Dr. W. Litaker of the Biotechnology Center of University of North Carolina for his valuable discussions, and Drs. A. Periasamy, G. Gordon and P. Wodnicki for their expert assistance in using computer for DNA database analysis. This research was supported by grant AG07218 from NIH to B.H.",

year = "1995",

doi = "10.1016/0378-1119(95)00542-E",

language = "English (US)",

volume = "165",

pages = "267--271",

journal = "Gene",

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TY - JOUR

T1 - Cloning of a rat cDNA encoding a novel LIM domain protein with high homology to rat RIL

AU - Wang, Haichao

AU - Harrison-Shostak, D. Corinne

AU - Lemasters, John J.

AU - Herman, Brian

N1 - Funding Information: We thank Dr. W. Litaker of the Biotechnology Center of University of North Carolina for his valuable discussions, and Drs. A. Periasamy, G. Gordon and P. Wodnicki for their expert assistance in using computer for DNA database analysis. This research was supported by grant AG07218 from NIH to B.H.

PY - 1995

Y1 - 1995

N2 - A complementary DNA (1392 bp) encoding a protein with high homology to rat reversion-induced LIM (RIL) protein was cloned from rat hepatocytes by differential screening of a subtractive (normoxic minus hypoxic) λGEM-2 cDNA library. This cDNA clone, denoted CLP-36, encodes a 327-amino-acid (aa) protein that contained a restrictively conserved LIM (a Cys-rich domain with consensus aa sequence C-X2-C-X17-19-H-X2-C-X2-C-X2-C-X16-20-C-X2-C/ H/D that was initially identified in homeodomain proteins, Lin-11 [Freyd et al., Nature 344 (1990) 876-879], Isl-1 [Karlsson et al., Nature 344 (1990) 879-882] and Mec-3 [Way et al., Cell 54 (1988) 5-16] in the C-terminal portion (aa 192-327). It shared an overall 45.1% identity to a rat LIM domain RIL protein [Kiess et al., Oncogene 10 (1995) 61-68], with 62.0% identity in the N terminus (aa 1-89) and 50.0% identity in the C terminus (aa 188-327). The rat CLP-36 mRNA is expressed most abundantly in heart, lung and liver, moderately in spleen and skeletal muscle, and at extremely low levels (if at all) in testis and brain tissues. Northern blotting analysis of total RNA extracted from normoxic or hypoxic rat hepatocytes, with a fragment of clone CLP-36 as probe, demonstrated a single band with a mobility corresponding to a size of 1.4 kb, whose level was significantly decreased during chemical hypoxia.

AB - A complementary DNA (1392 bp) encoding a protein with high homology to rat reversion-induced LIM (RIL) protein was cloned from rat hepatocytes by differential screening of a subtractive (normoxic minus hypoxic) λGEM-2 cDNA library. This cDNA clone, denoted CLP-36, encodes a 327-amino-acid (aa) protein that contained a restrictively conserved LIM (a Cys-rich domain with consensus aa sequence C-X2-C-X17-19-H-X2-C-X2-C-X2-C-X16-20-C-X2-C/ H/D that was initially identified in homeodomain proteins, Lin-11 [Freyd et al., Nature 344 (1990) 876-879], Isl-1 [Karlsson et al., Nature 344 (1990) 879-882] and Mec-3 [Way et al., Cell 54 (1988) 5-16] in the C-terminal portion (aa 192-327). It shared an overall 45.1% identity to a rat LIM domain RIL protein [Kiess et al., Oncogene 10 (1995) 61-68], with 62.0% identity in the N terminus (aa 1-89) and 50.0% identity in the C terminus (aa 188-327). The rat CLP-36 mRNA is expressed most abundantly in heart, lung and liver, moderately in spleen and skeletal muscle, and at extremely low levels (if at all) in testis and brain tissues. Northern blotting analysis of total RNA extracted from normoxic or hypoxic rat hepatocytes, with a fragment of clone CLP-36 as probe, demonstrated a single band with a mobility corresponding to a size of 1.4 kb, whose level was significantly decreased during chemical hypoxia.

KW - Hepatocyte

KW - PCR

KW - RACE

KW - amino-acid sequence

KW - chemical hypoxia

KW - nucleotide sequence

KW - phage library

KW - recombinant DNA

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U2 - 10.1016/0378-1119(95)00542-E

DO - 10.1016/0378-1119(95)00542-E

M3 - Article

C2 - 8522188

AN - SCOPUS:0028800393

SN - 0378-1119

VL - 165

SP - 267

EP - 271

JO - Gene

JF - Gene

IS - 2

ER -

Cloning of a rat cDNA encoding a novel LIM domain protein with high homology to rat RIL

Abstract

Bibliographical note

Keywords

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