On the Origin and Evolution of the Mosquito Male-determining Factor Nix

James K. Biedler; Azadeh Aryan; Yumin Qi; Aihua Wang; Ellen O. Martinson; Daniel A. Hartman; Fan Yang; Atashi Sharma; Katherine S. Morton; Mark Potters; Chujia Chen; Stephen L. Dobson; Gregory D. Ebel; Rebekah C. Kading; Sally Paulson; Rui De Xue; Michael R. Strand; Zhijian Tu

doi:10.1093/molbev/msad276

On the Origin and Evolution of the Mosquito Male-determining Factor Nix

James K. Biedler, Azadeh Aryan, Yumin Qi, Aihua Wang, Ellen O. Martinson, Daniel A. Hartman, Fan Yang, Atashi Sharma, Katherine S. Morton, Mark Potters, Chujia Chen, Stephen L. Dobson, Gregory D. Ebel, Rebekah C. Kading, Sally Paulson, Rui De Xue, Michael R. Strand, Zhijian Tu

Entomology

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

4 Scopus citations

Abstract

The mosquito family Culicidae is divided into 2 subfamilies named the Culicinae and Anophelinae. Nix, the dominant male-determining factor, has only been found in the culicines Aedes aegypti and Aedes albopictus, 2 important arboviral vectors that belong to the subgenus Stegomyia. Here we performed sex-specific whole-genome sequencing and RNAseq of divergent mosquito species and explored additional male-inclusive datasets to investigate the distribution of Nix. Except for the Culex genus, Nix homologs were found in all species surveyed from the Culicinae subfamily, including 12 additional species from 3 highly divergent tribes comprising 4 genera, suggesting Nix originated at least 133 to 165 million years ago (MYA). Heterologous expression of 1 of 3 divergent Nix open reading frames (ORFs) in Ae. aegypti resulted in partial masculinization of genetic females as evidenced by morphology and doublesex splicing. Phylogenetic analysis suggests Nix is related to femaleless (fle), a recently described intermediate sex-determining factor found exclusively in anopheline mosquitoes. Nix from all species has a conserved structure, including 3 RNA-recognition motifs (RRMs), as does fle. However, Nix has evolved at a much faster rate than fle. The RRM3 of both Nix and fle are distantly related to the single RRM of a widely distributed and conserved splicing factor transformer-2 (tra2). The RRM3-based phylogenetic analysis suggests this domain in Nix and fle may have evolved from tra2 or a tra2-related gene in a common ancestor of mosquitoes. Our results provide insights into the evolution of sex determination in mosquitoes and will inform broad applications of mosquito-control strategies based on manipulating sex ratios toward nonbiting males.

Original language	English
Article number	msad276
Journal	Molecular Biology and Evolution
Volume	41
Issue number	1
DOIs	https://doi.org/10.1093/molbev/msad276
State	Published - Jan 1 2024

Bibliographical note

Publisher Copyright:
© The Author(s) 2023. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

Funding

This work is supported by National Institutes of Health grants R01AI123338, 1R01AI157491 and R01AI179056 and the Virginia Agriculture Experimental Station. We thank Jeff Powell at Yale University and Daniel Dixon at the Anastasia Mosquito Control District (St. Augustine, FL, USA) for kindly providing the Aedes mascarensis and Psorophora columbiae samples. This work is supported by National Institutes of Health grants R01AI123338, 1R01AI157491 and R01AI179056 and the Virginia Agriculture Experimental Station. We thank Jeff Powell at Yale University and Daniel Dixon at the Anastasia Mosquito Control District (St. Augustine, FL, USA) for kindly providing the Aedes mascarensis and Psorophora columbiae samples.

Funders	Funder number
Daniel Dixon
National Institutes of Health (NIH)	1R01AI157491, R01AI123338, R01AI179056
National Institutes of Health (NIH)
Yale University
Virginia Agricultural Experiment Station, Virginia Polytechnic Institute and State University

Keywords

genetic engineering
homomorphic sex chromosome
male-determining factor
mosquito control
sex determination

ASJC Scopus subject areas

Ecology, Evolution, Behavior and Systematics
Molecular Biology
Genetics

Access to Document

10.1093/molbev/msad276

Cite this

Biedler, J. K., Aryan, A., Qi, Y., Wang, A., Martinson, E. O., Hartman, D. A., Yang, F., Sharma, A., Morton, K. S., Potters, M., Chen, C., Dobson, S. L., Ebel, G. D., Kading, R. C., Paulson, S., Xue, R. D., Strand, M. R., & Tu, Z. (2024). On the Origin and Evolution of the Mosquito Male-determining Factor Nix. Molecular Biology and Evolution, 41(1), Article msad276. https://doi.org/10.1093/molbev/msad276

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title = "On the Origin and Evolution of the Mosquito Male-determining Factor Nix",

abstract = "The mosquito family Culicidae is divided into 2 subfamilies named the Culicinae and Anophelinae. Nix, the dominant male-determining factor, has only been found in the culicines Aedes aegypti and Aedes albopictus, 2 important arboviral vectors that belong to the subgenus Stegomyia. Here we performed sex-specific whole-genome sequencing and RNAseq of divergent mosquito species and explored additional male-inclusive datasets to investigate the distribution of Nix. Except for the Culex genus, Nix homologs were found in all species surveyed from the Culicinae subfamily, including 12 additional species from 3 highly divergent tribes comprising 4 genera, suggesting Nix originated at least 133 to 165 million years ago (MYA). Heterologous expression of 1 of 3 divergent Nix open reading frames (ORFs) in Ae. aegypti resulted in partial masculinization of genetic females as evidenced by morphology and doublesex splicing. Phylogenetic analysis suggests Nix is related to femaleless (fle), a recently described intermediate sex-determining factor found exclusively in anopheline mosquitoes. Nix from all species has a conserved structure, including 3 RNA-recognition motifs (RRMs), as does fle. However, Nix has evolved at a much faster rate than fle. The RRM3 of both Nix and fle are distantly related to the single RRM of a widely distributed and conserved splicing factor transformer-2 (tra2). The RRM3-based phylogenetic analysis suggests this domain in Nix and fle may have evolved from tra2 or a tra2-related gene in a common ancestor of mosquitoes. Our results provide insights into the evolution of sex determination in mosquitoes and will inform broad applications of mosquito-control strategies based on manipulating sex ratios toward nonbiting males.",

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author = "Biedler, \{James K.\} and Azadeh Aryan and Yumin Qi and Aihua Wang and Martinson, \{Ellen O.\} and Hartman, \{Daniel A.\} and Fan Yang and Atashi Sharma and Morton, \{Katherine S.\} and Mark Potters and Chujia Chen and Dobson, \{Stephen L.\} and Ebel, \{Gregory D.\} and Kading, \{Rebekah C.\} and Sally Paulson and Xue, \{Rui De\} and Strand, \{Michael R.\} and Zhijian Tu",

note = "Publisher Copyright: {\textcopyright} The Author(s) 2023. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.",

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Biedler, JK, Aryan, A, Qi, Y, Wang, A, Martinson, EO, Hartman, DA, Yang, F, Sharma, A, Morton, KS, Potters, M, Chen, C, Dobson, SL, Ebel, GD, Kading, RC, Paulson, S, Xue, RD, Strand, MR & Tu, Z 2024, 'On the Origin and Evolution of the Mosquito Male-determining Factor Nix', Molecular Biology and Evolution, vol. 41, no. 1, msad276. https://doi.org/10.1093/molbev/msad276

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T1 - On the Origin and Evolution of the Mosquito Male-determining Factor Nix

AU - Biedler, James K.

AU - Aryan, Azadeh

AU - Qi, Yumin

AU - Wang, Aihua

AU - Martinson, Ellen O.

AU - Hartman, Daniel A.

AU - Yang, Fan

AU - Sharma, Atashi

AU - Morton, Katherine S.

AU - Potters, Mark

AU - Chen, Chujia

AU - Dobson, Stephen L.

AU - Ebel, Gregory D.

AU - Kading, Rebekah C.

AU - Paulson, Sally

AU - Xue, Rui De

AU - Strand, Michael R.

AU - Tu, Zhijian

N1 - Publisher Copyright: © The Author(s) 2023. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

PY - 2024/1/1

Y1 - 2024/1/1

N2 - The mosquito family Culicidae is divided into 2 subfamilies named the Culicinae and Anophelinae. Nix, the dominant male-determining factor, has only been found in the culicines Aedes aegypti and Aedes albopictus, 2 important arboviral vectors that belong to the subgenus Stegomyia. Here we performed sex-specific whole-genome sequencing and RNAseq of divergent mosquito species and explored additional male-inclusive datasets to investigate the distribution of Nix. Except for the Culex genus, Nix homologs were found in all species surveyed from the Culicinae subfamily, including 12 additional species from 3 highly divergent tribes comprising 4 genera, suggesting Nix originated at least 133 to 165 million years ago (MYA). Heterologous expression of 1 of 3 divergent Nix open reading frames (ORFs) in Ae. aegypti resulted in partial masculinization of genetic females as evidenced by morphology and doublesex splicing. Phylogenetic analysis suggests Nix is related to femaleless (fle), a recently described intermediate sex-determining factor found exclusively in anopheline mosquitoes. Nix from all species has a conserved structure, including 3 RNA-recognition motifs (RRMs), as does fle. However, Nix has evolved at a much faster rate than fle. The RRM3 of both Nix and fle are distantly related to the single RRM of a widely distributed and conserved splicing factor transformer-2 (tra2). The RRM3-based phylogenetic analysis suggests this domain in Nix and fle may have evolved from tra2 or a tra2-related gene in a common ancestor of mosquitoes. Our results provide insights into the evolution of sex determination in mosquitoes and will inform broad applications of mosquito-control strategies based on manipulating sex ratios toward nonbiting males.

AB - The mosquito family Culicidae is divided into 2 subfamilies named the Culicinae and Anophelinae. Nix, the dominant male-determining factor, has only been found in the culicines Aedes aegypti and Aedes albopictus, 2 important arboviral vectors that belong to the subgenus Stegomyia. Here we performed sex-specific whole-genome sequencing and RNAseq of divergent mosquito species and explored additional male-inclusive datasets to investigate the distribution of Nix. Except for the Culex genus, Nix homologs were found in all species surveyed from the Culicinae subfamily, including 12 additional species from 3 highly divergent tribes comprising 4 genera, suggesting Nix originated at least 133 to 165 million years ago (MYA). Heterologous expression of 1 of 3 divergent Nix open reading frames (ORFs) in Ae. aegypti resulted in partial masculinization of genetic females as evidenced by morphology and doublesex splicing. Phylogenetic analysis suggests Nix is related to femaleless (fle), a recently described intermediate sex-determining factor found exclusively in anopheline mosquitoes. Nix from all species has a conserved structure, including 3 RNA-recognition motifs (RRMs), as does fle. However, Nix has evolved at a much faster rate than fle. The RRM3 of both Nix and fle are distantly related to the single RRM of a widely distributed and conserved splicing factor transformer-2 (tra2). The RRM3-based phylogenetic analysis suggests this domain in Nix and fle may have evolved from tra2 or a tra2-related gene in a common ancestor of mosquitoes. Our results provide insights into the evolution of sex determination in mosquitoes and will inform broad applications of mosquito-control strategies based on manipulating sex ratios toward nonbiting males.

KW - genetic engineering

KW - homomorphic sex chromosome

KW - male-determining factor

KW - mosquito control

KW - sex determination

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ER -

On the Origin and Evolution of the Mosquito Male-determining Factor Nix

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

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